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Influence of added salt on the measured distribution of the radius a p and refractive index n p of <t>BSA-PAH</t> complexes. Each point in the scatter plots represents the properties of a single aggregate and is colored by the relative density of observations, ρ ( a p , n p ). (a) BSA complexed with PAH in Tris buffer (1100 aggregates). (b) The same sample with 0.1 M NaCl (1200 aggregates). (c) and (d) present the projected relative size distributions, ρ ( a p ), from (a) and (b), respectively. Shaded regions represent the instrumental and statistical error.
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Images

1) Product Images from "Holographic Characterization of Protein Aggregates"

Article Title: Holographic Characterization of Protein Aggregates

Journal: Journal of pharmaceutical sciences

doi: 10.1016/j.xphs.2015.12.018

Influence of added salt on the measured distribution of the radius a p and refractive index n p of BSA-PAH complexes. Each point in the scatter plots represents the properties of a single aggregate and is colored by the relative density of observations, ρ ( a p , n p ). (a) BSA complexed with PAH in Tris buffer (1100 aggregates). (b) The same sample with 0.1 M NaCl (1200 aggregates). (c) and (d) present the projected relative size distributions, ρ ( a p ), from (a) and (b), respectively. Shaded regions represent the instrumental and statistical error.
Figure Legend Snippet: Influence of added salt on the measured distribution of the radius a p and refractive index n p of BSA-PAH complexes. Each point in the scatter plots represents the properties of a single aggregate and is colored by the relative density of observations, ρ ( a p , n p ). (a) BSA complexed with PAH in Tris buffer (1100 aggregates). (b) The same sample with 0.1 M NaCl (1200 aggregates). (c) and (d) present the projected relative size distributions, ρ ( a p ), from (a) and (b), respectively. Shaded regions represent the instrumental and statistical error.

Techniques Used:

Influence of aggregate morphology on holographic characterization. Holograms of typical aggregates arranged in order of increasing discrepancy between measured and fit holograms. Column (a) shows 160 pixel × 160 pixel regions of interest from the microscope’s field of view, centered on features automatically identified as candidate BSA-PAH complexes. Column (b) shows fits to the Lorenz-Mie theory for holograms formed by spheres. Column (c) shows radial profiles of the experimental hologram (black curves) overlaid with the radial profile of the fits (red curves). Shaded regions represent the estimated experimental uncertainty. Column (d) shows Rayleigh-Sommerfeld reconstructions of the aggregates’ 3-dimensional structures obtained from the experimental holograms. Grayscale images are projections of the reconstructions, which resemble equivalent bright-field images at optimal focus. Superimposed circles indicate fit estimates for the particle size.
Figure Legend Snippet: Influence of aggregate morphology on holographic characterization. Holograms of typical aggregates arranged in order of increasing discrepancy between measured and fit holograms. Column (a) shows 160 pixel × 160 pixel regions of interest from the microscope’s field of view, centered on features automatically identified as candidate BSA-PAH complexes. Column (b) shows fits to the Lorenz-Mie theory for holograms formed by spheres. Column (c) shows radial profiles of the experimental hologram (black curves) overlaid with the radial profile of the fits (red curves). Shaded regions represent the estimated experimental uncertainty. Column (d) shows Rayleigh-Sommerfeld reconstructions of the aggregates’ 3-dimensional structures obtained from the experimental holograms. Grayscale images are projections of the reconstructions, which resemble equivalent bright-field images at optimal focus. Superimposed circles indicate fit estimates for the particle size.

Techniques Used: Microscopy

Characterization of BSA-PAH complexes by dynamic light scattering (DLS). Values represent the percentage, P ( a h ), of the scattered light’s intensity due to scatterers of a given hydrodynamic radius, a h . The arrow indicates a small peak in both distributions around a h = 2.8μm.
Figure Legend Snippet: Characterization of BSA-PAH complexes by dynamic light scattering (DLS). Values represent the percentage, P ( a h ), of the scattered light’s intensity due to scatterers of a given hydrodynamic radius, a h . The arrow indicates a small peak in both distributions around a h = 2.8μm.

Techniques Used:

2) Product Images from "Potential of electrospun cationic BSA fibers to guide osteogenic MSC differentiation via surface charge and fibrous topography"

Article Title: Potential of electrospun cationic BSA fibers to guide osteogenic MSC differentiation via surface charge and fibrous topography

Journal: Scientific Reports

doi: 10.1038/s41598-019-56508-6

Analysis of adhesion of MSCs on cBSA- and BSA-fibers. ( A ) Bar graphs of the metabolic activity of MSCs on cBSA- and BSA-fibers and TCP. Error bars represent SE. N = 3 independent experiments. Student’s t-test was used to assess potential statistical differences. ( B ) SEM images of MSCs (violet) on cBSA- (left) and BSA-fibers (right) colored in light red. Scale bar: 20 µm. ( C ) Fluorescence microscopy images from MSCs on cBSA-fibers, BSA-fibers, cBSA-coated glass, BSA-coated glass and untreated glass. F-actin of the cytoskeleton is stained in green with phalloidin Alexa flour 488 and the nuclei are stained in blue (DAPI). Scale bar: 50 µm. Schematic drawing of cells morphology on different samples created with the Matlab plugin phenoplot (lower row). The legend of schematic drawings is shown left. N = 3 independent experiments. Bar graphs of the values included in the phenoplot are included as supplementary information in Fig. S3 . ( D ) Immunofluorescence staining from MSCs on cBSA- and BSA-fibers as well as cBSA- and BSA-coated glass and untreated glass. The nuclei are stained blue (DAPI), actin filaments are stained in green and vinculin is marked in red. Top row shows an overlay picture of all channels and the lower rows show vinculin and actin staining alone. Scale bar: 10 µm. ( E ) Bar graph of quantified length of FACs in cells on different surfaces (cBSA-fibers, BSA-fibers, cBSA-coated glass, BSA-coated glass and glass) (left). Up to 10 FACs per cell were measured and the mean of the FACs from 36–60 cells with visible FACs from two different donors is plotted. Error bars represent the SD from 285–420 FACs. Statistical test: Anova with Tukey’s post-test. The percentage of cells with visible FACs (counted from 41–67 cells in total) on the different surfaces is plotted in the right bar graph. Fraction of cells displaying focal adhesions calculated from the sum of all assessed cells from N = 2 independent experiments.
Figure Legend Snippet: Analysis of adhesion of MSCs on cBSA- and BSA-fibers. ( A ) Bar graphs of the metabolic activity of MSCs on cBSA- and BSA-fibers and TCP. Error bars represent SE. N = 3 independent experiments. Student’s t-test was used to assess potential statistical differences. ( B ) SEM images of MSCs (violet) on cBSA- (left) and BSA-fibers (right) colored in light red. Scale bar: 20 µm. ( C ) Fluorescence microscopy images from MSCs on cBSA-fibers, BSA-fibers, cBSA-coated glass, BSA-coated glass and untreated glass. F-actin of the cytoskeleton is stained in green with phalloidin Alexa flour 488 and the nuclei are stained in blue (DAPI). Scale bar: 50 µm. Schematic drawing of cells morphology on different samples created with the Matlab plugin phenoplot (lower row). The legend of schematic drawings is shown left. N = 3 independent experiments. Bar graphs of the values included in the phenoplot are included as supplementary information in Fig. S3 . ( D ) Immunofluorescence staining from MSCs on cBSA- and BSA-fibers as well as cBSA- and BSA-coated glass and untreated glass. The nuclei are stained blue (DAPI), actin filaments are stained in green and vinculin is marked in red. Top row shows an overlay picture of all channels and the lower rows show vinculin and actin staining alone. Scale bar: 10 µm. ( E ) Bar graph of quantified length of FACs in cells on different surfaces (cBSA-fibers, BSA-fibers, cBSA-coated glass, BSA-coated glass and glass) (left). Up to 10 FACs per cell were measured and the mean of the FACs from 36–60 cells with visible FACs from two different donors is plotted. Error bars represent the SD from 285–420 FACs. Statistical test: Anova with Tukey’s post-test. The percentage of cells with visible FACs (counted from 41–67 cells in total) on the different surfaces is plotted in the right bar graph. Fraction of cells displaying focal adhesions calculated from the sum of all assessed cells from N = 2 independent experiments.

Techniques Used: Activity Assay, Fluorescence, Microscopy, Staining, Immunofluorescence, FACS

3) Product Images from "Dectin-2-Targeted Antifungal Liposomes Exhibit Enhanced Efficacy"

Article Title: Dectin-2-Targeted Antifungal Liposomes Exhibit Enhanced Efficacy

Journal: mSphere

doi: 10.1128/mSphere.00715-19

sDectin-2-coated amphotericin B-loaded liposome inhibition and killing of C. albicans , C. neoformans , and A. fumigatus . (A) C. albicans with DEC2-AmB-LLs. Cells in the pseudohyphal and early hyphal stage grown in RPMI medium plus 0.5% BSA on 96-well polystyrene microtiter plates. Cells were treated for 30 min with liposomes delivering 1.0., 0.5, 0.25, and 0.12 μM AmB to the medium as indicated, washed twice with medium, grown for an additional 16 h, and then assayed for metabolic activity using CellTiter-Blue (CTB) reagent. (B) C. neoformans with DEC2-AmB-LLs. C. neoformans cells were grown in liquid YPD medium plus 0.5% BSA with vigorous shaking for 2 h and treated for 4 h or overnight with liposomes delivering 0.4, 0.2, or 0.1 μM AmB to the medium as indicated. Cells were diluted and plated on YPD medium, and CFU were counted from multiple plates. (C) A. fumigatus with DEC2-AmB-LLs. Conidia were germinated for 9 h in VMM plus glucose plus 0.5% BSA in 96-well polystyrene microtiter plates, treated for 2 h with liposomes delivering 0.5 and 0.25 μM AmB to the medium as indicated, washed twice with medium, grown overnight, and then assayed for metabolic activity using CTB reagent in RPMI lacking phenol red indicator plus 0.5% BSA. The control wells were overgrown with hyphae protruding out the medium. (D) A. fumigatus with DEC1-AmB-LLs. Assay conditions were similar to the assay in panel C, except liposomes were first diluted into LDB1 buffer (PBS plus 0.5%BSA plus 1 mM BME) prior to dilution into growth medium. For CTB assays in panels A, C, and D, the fluorescence background from medium incubated with CTB reagent was subtracted. Standard errors are shown for all values, and fold differences and P values were estimated comparing the performance of AmB-LLs to DEC2-AmB-LLs. Two or more biological replicates gave similar results.
Figure Legend Snippet: sDectin-2-coated amphotericin B-loaded liposome inhibition and killing of C. albicans , C. neoformans , and A. fumigatus . (A) C. albicans with DEC2-AmB-LLs. Cells in the pseudohyphal and early hyphal stage grown in RPMI medium plus 0.5% BSA on 96-well polystyrene microtiter plates. Cells were treated for 30 min with liposomes delivering 1.0., 0.5, 0.25, and 0.12 μM AmB to the medium as indicated, washed twice with medium, grown for an additional 16 h, and then assayed for metabolic activity using CellTiter-Blue (CTB) reagent. (B) C. neoformans with DEC2-AmB-LLs. C. neoformans cells were grown in liquid YPD medium plus 0.5% BSA with vigorous shaking for 2 h and treated for 4 h or overnight with liposomes delivering 0.4, 0.2, or 0.1 μM AmB to the medium as indicated. Cells were diluted and plated on YPD medium, and CFU were counted from multiple plates. (C) A. fumigatus with DEC2-AmB-LLs. Conidia were germinated for 9 h in VMM plus glucose plus 0.5% BSA in 96-well polystyrene microtiter plates, treated for 2 h with liposomes delivering 0.5 and 0.25 μM AmB to the medium as indicated, washed twice with medium, grown overnight, and then assayed for metabolic activity using CTB reagent in RPMI lacking phenol red indicator plus 0.5% BSA. The control wells were overgrown with hyphae protruding out the medium. (D) A. fumigatus with DEC1-AmB-LLs. Assay conditions were similar to the assay in panel C, except liposomes were first diluted into LDB1 buffer (PBS plus 0.5%BSA plus 1 mM BME) prior to dilution into growth medium. For CTB assays in panels A, C, and D, the fluorescence background from medium incubated with CTB reagent was subtracted. Standard errors are shown for all values, and fold differences and P values were estimated comparing the performance of AmB-LLs to DEC2-AmB-LLs. Two or more biological replicates gave similar results.

Techniques Used: Inhibition, Activity Assay, CtB Assay, Fluorescence, Incubation

4) Product Images from "TNFα/IFNγ Mediated Intestinal Epithelial Barrier Dysfunction Is Attenuated by MicroRNA-93 Downregulation of PTK6 in Mouse Colonic Epithelial Cells"

Article Title: TNFα/IFNγ Mediated Intestinal Epithelial Barrier Dysfunction Is Attenuated by MicroRNA-93 Downregulation of PTK6 in Mouse Colonic Epithelial Cells

Journal: PLoS ONE

doi: 10.1371/journal.pone.0154351

PTK6 contributes to the cytokine cocktail TNFα/IFNγ induced barrier dysfunction and FoxO1 nuclear translocation in YAMC. Post-confluent, mortalized WT or PTK6-/- YAMC monolayers were treated with vehicle control (0.1% BSA in PBS) or TNFα (100ng/ml) and IFNγ (500U/ml) 16 hours. A) Nuclei (blue), ZO-1 (green), and FoxO1 (red) were detected via confocal microscopy. B) Quantitation of the ratio of ZO-1 intensity in the cytoplasm over cell-cell junction from Fig 3A. C) Quantitation for nuclear FoxO1 intensity of FoxO1 stain in Fig 3A. D) Western blot indicating expression levels of ZO-1 in total cell lysates (please see S5 Fig for full blot scan). *p
Figure Legend Snippet: PTK6 contributes to the cytokine cocktail TNFα/IFNγ induced barrier dysfunction and FoxO1 nuclear translocation in YAMC. Post-confluent, mortalized WT or PTK6-/- YAMC monolayers were treated with vehicle control (0.1% BSA in PBS) or TNFα (100ng/ml) and IFNγ (500U/ml) 16 hours. A) Nuclei (blue), ZO-1 (green), and FoxO1 (red) were detected via confocal microscopy. B) Quantitation of the ratio of ZO-1 intensity in the cytoplasm over cell-cell junction from Fig 3A. C) Quantitation for nuclear FoxO1 intensity of FoxO1 stain in Fig 3A. D) Western blot indicating expression levels of ZO-1 in total cell lysates (please see S5 Fig for full blot scan). *p

Techniques Used: Translocation Assay, Confocal Microscopy, Quantitation Assay, Staining, Western Blot, Expressing

TNFα/IFNγ mediated barrier dysfunction involves activation of PTK6, JNK, and downregulation of claudin-3 in YAMC. A) Mortalized, 2-day post-confluent YAMC monolayers were treated with vehicle control (0.1% BSA in PBS) or TNFα (100ng/ml) and IFNγ (500U/ml) at the indicated time points then assayed for changes in expression of the indicated proteins (please refer to S3 Fig for full blot scans). B) RNA was isolated then quantitated for PTK6 levels with qPCR. Error bars represent standard error for 3 separate experiments (*p
Figure Legend Snippet: TNFα/IFNγ mediated barrier dysfunction involves activation of PTK6, JNK, and downregulation of claudin-3 in YAMC. A) Mortalized, 2-day post-confluent YAMC monolayers were treated with vehicle control (0.1% BSA in PBS) or TNFα (100ng/ml) and IFNγ (500U/ml) at the indicated time points then assayed for changes in expression of the indicated proteins (please refer to S3 Fig for full blot scans). B) RNA was isolated then quantitated for PTK6 levels with qPCR. Error bars represent standard error for 3 separate experiments (*p

Techniques Used: Activation Assay, Expressing, Isolation, Real-time Polymerase Chain Reaction

PTK6 contributes to the cytokine cocktail TNFα/IFNγ induced barrier dysfunction and FoxO1 nuclear translocation in YAMC. Post-confluent, mortalized WT or PTK6-/- YAMC monolayers were treated with vehicle control (0.1% BSA in PBS) or TNFα (100ng/ml) and IFNγ (500U/ml) 16 hours. A) Nuclei (blue), ZO-1 (green), and FoxO1 (red) were detected via confocal microscopy. B) Quantitation of the ratio of ZO-1 intensity in the cytoplasm over cell-cell junction from Fig 3A. C) Quantitation for nuclear FoxO1 intensity of FoxO1 stain in Fig 3A. D) Western blot indicating expression levels of ZO-1 in total cell lysates (please see S5 Fig for full blot scan). *p
Figure Legend Snippet: PTK6 contributes to the cytokine cocktail TNFα/IFNγ induced barrier dysfunction and FoxO1 nuclear translocation in YAMC. Post-confluent, mortalized WT or PTK6-/- YAMC monolayers were treated with vehicle control (0.1% BSA in PBS) or TNFα (100ng/ml) and IFNγ (500U/ml) 16 hours. A) Nuclei (blue), ZO-1 (green), and FoxO1 (red) were detected via confocal microscopy. B) Quantitation of the ratio of ZO-1 intensity in the cytoplasm over cell-cell junction from Fig 3A. C) Quantitation for nuclear FoxO1 intensity of FoxO1 stain in Fig 3A. D) Western blot indicating expression levels of ZO-1 in total cell lysates (please see S5 Fig for full blot scan). *p

Techniques Used: Translocation Assay, Confocal Microscopy, Quantitation Assay, Staining, Western Blot, Expressing

5) Product Images from "Oligosaccharide conjugates of Bordetella pertussis and bronchiseptica induce bactericidal antibodies, an addition to pertussis vaccine"

Article Title: Oligosaccharide conjugates of Bordetella pertussis and bronchiseptica induce bactericidal antibodies, an addition to pertussis vaccine

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1100782108

SDS/PAGE and Western blots of B. pertussis Tohama I (lane 1), B. bronchiseptica RBwbmΔ (lane 2), B. bronchiseptica RBA2b (lane 3), and B. bronchiseptica RBB1a (lane 4) LPS. ( A ) SDS/PAGE. ( B – D ) Western blots with anti- B. pertussis Tohama I bacteria (one band A trisaccharide repeat; B ) with anti-BSA/RBA2b-F1 (four repeats of band A trisaccharide; C ; similar picture was obtained with anti-BSA/RBA2b-F2 and BSA/RBA2b-F3 conjugates with three and two repeats of band A trisaccharide); and with anti-BSA/ RBA2b-F4 (one repeat of band A trisaccharide; D ).
Figure Legend Snippet: SDS/PAGE and Western blots of B. pertussis Tohama I (lane 1), B. bronchiseptica RBwbmΔ (lane 2), B. bronchiseptica RBA2b (lane 3), and B. bronchiseptica RBB1a (lane 4) LPS. ( A ) SDS/PAGE. ( B – D ) Western blots with anti- B. pertussis Tohama I bacteria (one band A trisaccharide repeat; B ) with anti-BSA/RBA2b-F1 (four repeats of band A trisaccharide; C ; similar picture was obtained with anti-BSA/RBA2b-F2 and BSA/RBA2b-F3 conjugates with three and two repeats of band A trisaccharide); and with anti-BSA/ RBA2b-F4 (one repeat of band A trisaccharide; D ).

Techniques Used: SDS Page, Western Blot

6) Product Images from "TNFα/IFNγ Mediated Intestinal Epithelial Barrier Dysfunction Is Attenuated by MicroRNA-93 Downregulation of PTK6 in Mouse Colonic Epithelial Cells"

Article Title: TNFα/IFNγ Mediated Intestinal Epithelial Barrier Dysfunction Is Attenuated by MicroRNA-93 Downregulation of PTK6 in Mouse Colonic Epithelial Cells

Journal: PLoS ONE

doi: 10.1371/journal.pone.0154351

PTK6 contributes to the cytokine cocktail TNFα/IFNγ induced barrier dysfunction and FoxO1 nuclear translocation in YAMC. Post-confluent, mortalized WT or PTK6-/- YAMC monolayers were treated with vehicle control (0.1% BSA in PBS) or TNFα (100ng/ml) and IFNγ (500U/ml) 16 hours. A) Nuclei (blue), ZO-1 (green), and FoxO1 (red) were detected via confocal microscopy. B) Quantitation of the ratio of ZO-1 intensity in the cytoplasm over cell-cell junction from Fig 3A. C) Quantitation for nuclear FoxO1 intensity of FoxO1 stain in Fig 3A. D) Western blot indicating expression levels of ZO-1 in total cell lysates (please see S5 Fig for full blot scan). *p
Figure Legend Snippet: PTK6 contributes to the cytokine cocktail TNFα/IFNγ induced barrier dysfunction and FoxO1 nuclear translocation in YAMC. Post-confluent, mortalized WT or PTK6-/- YAMC monolayers were treated with vehicle control (0.1% BSA in PBS) or TNFα (100ng/ml) and IFNγ (500U/ml) 16 hours. A) Nuclei (blue), ZO-1 (green), and FoxO1 (red) were detected via confocal microscopy. B) Quantitation of the ratio of ZO-1 intensity in the cytoplasm over cell-cell junction from Fig 3A. C) Quantitation for nuclear FoxO1 intensity of FoxO1 stain in Fig 3A. D) Western blot indicating expression levels of ZO-1 in total cell lysates (please see S5 Fig for full blot scan). *p

Techniques Used: Translocation Assay, Confocal Microscopy, Quantitation Assay, Staining, Western Blot, Expressing

TNFα/IFNγ mediated barrier dysfunction involves activation of PTK6, JNK, and downregulation of claudin-3 in YAMC. A) Mortalized, 2-day post-confluent YAMC monolayers were treated with vehicle control (0.1% BSA in PBS) or TNFα (100ng/ml) and IFNγ (500U/ml) at the indicated time points then assayed for changes in expression of the indicated proteins (please refer to S3 Fig for full blot scans). B) RNA was isolated then quantitated for PTK6 levels with qPCR. Error bars represent standard error for 3 separate experiments (*p
Figure Legend Snippet: TNFα/IFNγ mediated barrier dysfunction involves activation of PTK6, JNK, and downregulation of claudin-3 in YAMC. A) Mortalized, 2-day post-confluent YAMC monolayers were treated with vehicle control (0.1% BSA in PBS) or TNFα (100ng/ml) and IFNγ (500U/ml) at the indicated time points then assayed for changes in expression of the indicated proteins (please refer to S3 Fig for full blot scans). B) RNA was isolated then quantitated for PTK6 levels with qPCR. Error bars represent standard error for 3 separate experiments (*p

Techniques Used: Activation Assay, Expressing, Isolation, Real-time Polymerase Chain Reaction

PTK6 contributes to the cytokine cocktail TNFα/IFNγ induced barrier dysfunction and FoxO1 nuclear translocation in YAMC. Post-confluent, mortalized WT or PTK6-/- YAMC monolayers were treated with vehicle control (0.1% BSA in PBS) or TNFα (100ng/ml) and IFNγ (500U/ml) 16 hours. A) Nuclei (blue), ZO-1 (green), and FoxO1 (red) were detected via confocal microscopy. B) Quantitation of the ratio of ZO-1 intensity in the cytoplasm over cell-cell junction from Fig 3A. C) Quantitation for nuclear FoxO1 intensity of FoxO1 stain in Fig 3A. D) Western blot indicating expression levels of ZO-1 in total cell lysates (please see S5 Fig for full blot scan). *p
Figure Legend Snippet: PTK6 contributes to the cytokine cocktail TNFα/IFNγ induced barrier dysfunction and FoxO1 nuclear translocation in YAMC. Post-confluent, mortalized WT or PTK6-/- YAMC monolayers were treated with vehicle control (0.1% BSA in PBS) or TNFα (100ng/ml) and IFNγ (500U/ml) 16 hours. A) Nuclei (blue), ZO-1 (green), and FoxO1 (red) were detected via confocal microscopy. B) Quantitation of the ratio of ZO-1 intensity in the cytoplasm over cell-cell junction from Fig 3A. C) Quantitation for nuclear FoxO1 intensity of FoxO1 stain in Fig 3A. D) Western blot indicating expression levels of ZO-1 in total cell lysates (please see S5 Fig for full blot scan). *p

Techniques Used: Translocation Assay, Confocal Microscopy, Quantitation Assay, Staining, Western Blot, Expressing

7) Product Images from "Direct Cell Lysis for Single-Cell Gene Expression Profiling"

Article Title: Direct Cell Lysis for Single-Cell Gene Expression Profiling

Journal: Frontiers in Oncology

doi: 10.3389/fonc.2013.00274

Evaluation of direct cell lysis protocols . (A) The lysis yields of Gapdh , Vim , Dll1 , Jag1 , DNA, and RNA spike compared at 17 lysis conditions. Thirty-two astrocytes were sorted for each condition. Relative cDNA yields are presented as Cq-values on the left y -axis and relative transcript numbers on the right y -axis. The relative transcript number is expressed in percentage compared to the optimal lysis condition for each gene, assuming 100% RT efficiency and 100% PCR efficiency. Data are shown as mean ± SD ( n = 4). Missing data were excluded and are listed in Table S3 in Supplementary Material. (B) Mean cDNA yield of the transcripts. Expressions of Gapdh , Vim , Dll , and Jag1 were averaged and are compared to the overall optimal lysis condition (1 mg/ml BSA). Data are shown as mean ± SD ( n = 4). 7-deaz GTP, 7-deaza-2′ deoxyguanosine 5′ triphosphate lithium salt; GTC, guanidine thiocyanate; LPA, linear polyacrylamide; polyI, polyinosinic acid potassium salt; 2× RT buffer, 2× reverse transcription buffer; RT mix, 2× RT buffer, 5 μM random hexamers, 5 μM oligo-dT, and 1 mM dNTP.
Figure Legend Snippet: Evaluation of direct cell lysis protocols . (A) The lysis yields of Gapdh , Vim , Dll1 , Jag1 , DNA, and RNA spike compared at 17 lysis conditions. Thirty-two astrocytes were sorted for each condition. Relative cDNA yields are presented as Cq-values on the left y -axis and relative transcript numbers on the right y -axis. The relative transcript number is expressed in percentage compared to the optimal lysis condition for each gene, assuming 100% RT efficiency and 100% PCR efficiency. Data are shown as mean ± SD ( n = 4). Missing data were excluded and are listed in Table S3 in Supplementary Material. (B) Mean cDNA yield of the transcripts. Expressions of Gapdh , Vim , Dll , and Jag1 were averaged and are compared to the overall optimal lysis condition (1 mg/ml BSA). Data are shown as mean ± SD ( n = 4). 7-deaz GTP, 7-deaza-2′ deoxyguanosine 5′ triphosphate lithium salt; GTC, guanidine thiocyanate; LPA, linear polyacrylamide; polyI, polyinosinic acid potassium salt; 2× RT buffer, 2× reverse transcription buffer; RT mix, 2× RT buffer, 5 μM random hexamers, 5 μM oligo-dT, and 1 mM dNTP.

Techniques Used: Lysis, Polymerase Chain Reaction

mRNA accessibility over time . (A) mRNA accessibility over time in 1–4 mg/ml BSA, 50 ng/μl yeast tRNA, 1× RT buffer, and water. Five hundred astrocytes were lysed and kept in room temperature for 0, 1, 2, and 6 h. Cq-values are shown on the left y -axis and relative transcript numbers on the right y -axis. Relative transcript number is expressed in percentage compared to the 1 mg/ml BSA sample at 0 h, assuming 100% RT efficiency and 100% PCR efficiency. Data are shown as mean ± SD ( n = 4). (B) Mean RNA accessibility of the transcripts. Expression of Gapdh , Vim , Dll , and Jag1 were averaged and compared to the 1 mg/ml BSA condition at 0 h. (C) Percentage of positive data points. Missing data were excluded from subplots (A,B) and are shown in Table S5 in Supplementary Material. Four genes and four time points were analyzed per lysis condition. GTC, guanidine thiocyanate; 1× RT buffer, 1× reverse transcription buffer.
Figure Legend Snippet: mRNA accessibility over time . (A) mRNA accessibility over time in 1–4 mg/ml BSA, 50 ng/μl yeast tRNA, 1× RT buffer, and water. Five hundred astrocytes were lysed and kept in room temperature for 0, 1, 2, and 6 h. Cq-values are shown on the left y -axis and relative transcript numbers on the right y -axis. Relative transcript number is expressed in percentage compared to the 1 mg/ml BSA sample at 0 h, assuming 100% RT efficiency and 100% PCR efficiency. Data are shown as mean ± SD ( n = 4). (B) Mean RNA accessibility of the transcripts. Expression of Gapdh , Vim , Dll , and Jag1 were averaged and compared to the 1 mg/ml BSA condition at 0 h. (C) Percentage of positive data points. Missing data were excluded from subplots (A,B) and are shown in Table S5 in Supplementary Material. Four genes and four time points were analyzed per lysis condition. GTC, guanidine thiocyanate; 1× RT buffer, 1× reverse transcription buffer.

Techniques Used: Polymerase Chain Reaction, Expressing, Lysis

mRNA during freeze/thaw cycling . (A) Comparison of RNA accessibility after freeze/thaw cycles in 1–4 mg/ml BSA, 50 ng/μl yeast tRNA, 1× RT buffer and water. Five hundred astrocytes were lysed, frozen in −80°C and thawed in room temperature 1, 2, 3, or 6 times. Cq-values are shown on the left y -axis and relative transcript numbers on the right y -axis. Relative transcript number is expressed in percentage compared to the 1 mg/ml BSA sample thawed once, assuming 100% RT efficiency and 100% PCR efficiency. Data are shown as mean ± SD ( n = 4) (B) Mean RNA accessibility of the transcripts. Expression of Gapdh , Vim , Dll , and Jag1 were averaged and compared to the 1 mg/ml BSA sample thawed once. (C) Percentage of positive data points. Missing data were excluded from subplot (A,B) and are shown in Table S6 in Supplementary Material. Four genes and four different amounts of freeze/thaw cycles were analyzed per lysis condition. GTC, guanidine thiocyanate; 1× RT buffer, 1× reverse transcription buffer.
Figure Legend Snippet: mRNA during freeze/thaw cycling . (A) Comparison of RNA accessibility after freeze/thaw cycles in 1–4 mg/ml BSA, 50 ng/μl yeast tRNA, 1× RT buffer and water. Five hundred astrocytes were lysed, frozen in −80°C and thawed in room temperature 1, 2, 3, or 6 times. Cq-values are shown on the left y -axis and relative transcript numbers on the right y -axis. Relative transcript number is expressed in percentage compared to the 1 mg/ml BSA sample thawed once, assuming 100% RT efficiency and 100% PCR efficiency. Data are shown as mean ± SD ( n = 4) (B) Mean RNA accessibility of the transcripts. Expression of Gapdh , Vim , Dll , and Jag1 were averaged and compared to the 1 mg/ml BSA sample thawed once. (C) Percentage of positive data points. Missing data were excluded from subplot (A,B) and are shown in Table S6 in Supplementary Material. Four genes and four different amounts of freeze/thaw cycles were analyzed per lysis condition. GTC, guanidine thiocyanate; 1× RT buffer, 1× reverse transcription buffer.

Techniques Used: Polymerase Chain Reaction, Expressing, Lysis

Evaluation of direct cell lysis protocols . (A) The lysis yields of Gapdh , Vim , Dll1 , Jag1 , DNA, and RNA spike compared at 17 lysis conditions. Thirty-two astrocytes were sorted for each condition. Relative cDNA yields are presented as Cq-values on the left y -axis and relative transcript numbers on the right y -axis. The relative transcript number is expressed in percentage compared to the optimal lysis condition for each gene, assuming 100% RT efficiency and 100% PCR efficiency. Data are shown as mean ± SD ( n = 4). Missing data were excluded and are listed in Table S3 in Supplementary Material. (B) Mean cDNA yield of the transcripts. Expressions of Gapdh , Vim , Dll , and Jag1 were averaged and are compared to the overall optimal lysis condition (1 mg/ml BSA). Data are shown as mean ± SD ( n = 4). 7-deaz GTP, 7-deaza-2′ deoxyguanosine 5′ triphosphate lithium salt; GTC, guanidine thiocyanate; LPA, linear polyacrylamide; polyI, polyinosinic acid potassium salt; 2× RT buffer, 2× reverse transcription buffer; RT mix, 2× RT buffer, 5 μM random hexamers, 5 μM oligo-dT, and 1 mM dNTP.
Figure Legend Snippet: Evaluation of direct cell lysis protocols . (A) The lysis yields of Gapdh , Vim , Dll1 , Jag1 , DNA, and RNA spike compared at 17 lysis conditions. Thirty-two astrocytes were sorted for each condition. Relative cDNA yields are presented as Cq-values on the left y -axis and relative transcript numbers on the right y -axis. The relative transcript number is expressed in percentage compared to the optimal lysis condition for each gene, assuming 100% RT efficiency and 100% PCR efficiency. Data are shown as mean ± SD ( n = 4). Missing data were excluded and are listed in Table S3 in Supplementary Material. (B) Mean cDNA yield of the transcripts. Expressions of Gapdh , Vim , Dll , and Jag1 were averaged and are compared to the overall optimal lysis condition (1 mg/ml BSA). Data are shown as mean ± SD ( n = 4). 7-deaz GTP, 7-deaza-2′ deoxyguanosine 5′ triphosphate lithium salt; GTC, guanidine thiocyanate; LPA, linear polyacrylamide; polyI, polyinosinic acid potassium salt; 2× RT buffer, 2× reverse transcription buffer; RT mix, 2× RT buffer, 5 μM random hexamers, 5 μM oligo-dT, and 1 mM dNTP.

Techniques Used: Lysis, Polymerase Chain Reaction

8) Product Images from "Selective modulation of type 1 insulin-like growth factor receptor signaling and functions by ?1 integrins"

Article Title: Selective modulation of type 1 insulin-like growth factor receptor signaling and functions by ?1 integrins

Journal: The Journal of Cell Biology

doi: 10.1083/jcb.200403003

IGF binding to IGF-IR mediates adhesion of β 1C expressing cells to LN-1. (A) β 1A - and β 1C -CHO clones were transiently transfected with 486/STOP or vector alone. Cells were cultured in the absence of tet for 48 h. Cells were plated on either LN-1 or FN and incubated for 2 h at 37°C in the presence or in the absence of IGF-II. After incubation, cells were fixed and stained with β-gal. The number of attached cells transfected with 486/STOP cDNA was expressed as percentage of the number of attached cells transfected with vector alone, set at 100. (B and C) β 1C - and β 1A -PC3 clones were cultured in the absence of tet for 48 h. Cells were plated on BSA or LN-1 (B) or FN (C) at 37°C for 2 h in the presence or in the absence of IGF-I or α-IR3 or ni-mIgG. The experiments were repeated at least three times with similar results. Data are expressed as mean ± SEM.
Figure Legend Snippet: IGF binding to IGF-IR mediates adhesion of β 1C expressing cells to LN-1. (A) β 1A - and β 1C -CHO clones were transiently transfected with 486/STOP or vector alone. Cells were cultured in the absence of tet for 48 h. Cells were plated on either LN-1 or FN and incubated for 2 h at 37°C in the presence or in the absence of IGF-II. After incubation, cells were fixed and stained with β-gal. The number of attached cells transfected with 486/STOP cDNA was expressed as percentage of the number of attached cells transfected with vector alone, set at 100. (B and C) β 1C - and β 1A -PC3 clones were cultured in the absence of tet for 48 h. Cells were plated on BSA or LN-1 (B) or FN (C) at 37°C for 2 h in the presence or in the absence of IGF-I or α-IR3 or ni-mIgG. The experiments were repeated at least three times with similar results. Data are expressed as mean ± SEM.

Techniques Used: Binding Assay, Expressing, Clone Assay, Transfection, Plasmid Preparation, Cell Culture, Incubation, Staining

Down-regulation of β 1C expression in 267B1 cells inhibits IGF-I–stimulated cell adhesion to LN-1. (A) PrEC and PC3 cells were serum starved for 12 h, detached, and plated on LN-1 or FN at 37°C for 2 h in the presence or in the absence of IGF-I. Cell adhesion was analyzed by crystal violet staining. (B) pRNS-1-1 and 267B1 cells were lysed and immunoblotted with mAb to β 1C or normal IgM. (C) 267B1 cells were transiently transfected with pBJ1-RZ-β 1C or vector alone, lysed, and immunoblotted with mAb to β 1C , Ab to Akt, or Ab to IGF-IR. CHO-β 1C cell lysate was used as a positive control. (D) 267B1 cells were transiently transfected with pBJ1-RZ-β 1C or vector alone were detached and seeded on BSA or LN-1 or FN-coated plates at 37°C for 2 h in the presence or in the absence of IGF-I and stained with β-gal. Cell attachment was expressed as percentage of cells transfected with vector and attached to LN-1 in the absence of IGF-I, set at 100. Data are expressed as mean ± SEM. The experiments were repeated at least twice with similar results.
Figure Legend Snippet: Down-regulation of β 1C expression in 267B1 cells inhibits IGF-I–stimulated cell adhesion to LN-1. (A) PrEC and PC3 cells were serum starved for 12 h, detached, and plated on LN-1 or FN at 37°C for 2 h in the presence or in the absence of IGF-I. Cell adhesion was analyzed by crystal violet staining. (B) pRNS-1-1 and 267B1 cells were lysed and immunoblotted with mAb to β 1C or normal IgM. (C) 267B1 cells were transiently transfected with pBJ1-RZ-β 1C or vector alone, lysed, and immunoblotted with mAb to β 1C , Ab to Akt, or Ab to IGF-IR. CHO-β 1C cell lysate was used as a positive control. (D) 267B1 cells were transiently transfected with pBJ1-RZ-β 1C or vector alone were detached and seeded on BSA or LN-1 or FN-coated plates at 37°C for 2 h in the presence or in the absence of IGF-I and stained with β-gal. Cell attachment was expressed as percentage of cells transfected with vector and attached to LN-1 in the absence of IGF-I, set at 100. Data are expressed as mean ± SEM. The experiments were repeated at least twice with similar results.

Techniques Used: Expressing, Staining, Transfection, Plasmid Preparation, Positive Control, Cell Attachment Assay

IGF-IR and β 1C act synergistically to support cell adhesion to LN-1. (A) R− and R+ cells were transiently transfected with human β 1A or β 1C . Surface expression of β 1A or β 1C was analyzed by FACS ® using TS2/16 or 12CA5 as a negative control. Thick line, TS2/16; thin line, 12CA5. (B) R+ cells transiently transfected with β 1A or β 1C were detached and seeded on BSA or LN-1–coated plates at 37°C for 2 h in the presence or in the absence of IGF-II and stained with β-gal. (C) R− and R+ cells (10 6 ) transiently transfected with β 1A or β 1C were incubated with or without P4C10 (a-β 1 ) or 1C10 (neg-cont) on ice for 1 h. Cells were plated on BSA or LN-1 or FN at 37°C for 2 h and stained with β-gal. Attachment of cells transfected with β 1 -integrin cDNA was expressed as percentage of cells transfected with pBJ (B) or pBJ-β 1A (C) that were attached to LN-1, set at 100. The experiments were repeated at least twice with similar results. Data are expressed as mean ± SEM.
Figure Legend Snippet: IGF-IR and β 1C act synergistically to support cell adhesion to LN-1. (A) R− and R+ cells were transiently transfected with human β 1A or β 1C . Surface expression of β 1A or β 1C was analyzed by FACS ® using TS2/16 or 12CA5 as a negative control. Thick line, TS2/16; thin line, 12CA5. (B) R+ cells transiently transfected with β 1A or β 1C were detached and seeded on BSA or LN-1–coated plates at 37°C for 2 h in the presence or in the absence of IGF-II and stained with β-gal. (C) R− and R+ cells (10 6 ) transiently transfected with β 1A or β 1C were incubated with or without P4C10 (a-β 1 ) or 1C10 (neg-cont) on ice for 1 h. Cells were plated on BSA or LN-1 or FN at 37°C for 2 h and stained with β-gal. Attachment of cells transfected with β 1 -integrin cDNA was expressed as percentage of cells transfected with pBJ (B) or pBJ-β 1A (C) that were attached to LN-1, set at 100. The experiments were repeated at least twice with similar results. Data are expressed as mean ± SEM.

Techniques Used: Activated Clotting Time Assay, Transfection, Expressing, FACS, Negative Control, Staining, Incubation

β 1C enhances IGF-I– and II–mediated PC3 cell adhesion to LN-1. (A–D) β 1A - and β 1C -PC3 clones were cultured in the presence or in the absence of tet for 48 h. Cells were detached and plated on BSA or LN-1 or FN at 37°C for 2 h in the presence of IGF-II (A and B), of IGF-I (C and D), or of Ab to LN-1 (D) or ni-rIgG (D). In A, the differences in cell adhesion to LN-1 in the presence or in the absence of tet are statistically significant (*P ≤ 0.03). In C, the differences in cell adhesion to LN-1 between β 1A and β 1C expressing cells are statistically significant (*P ≤ 0.024). The experiments were repeated at least three times with similar results using two clones each of β 1A - and β 1C -PC3 cells. (E and F) β 1A - and β 1C -PC3 clones were cultured in the presence or in the absence of tet for 48 h. Cells were serum starved for 12 h, detached, and plated on LN-1 (E), FN or BSA (F) in serum-free medium at 37°C for 2 h in the presence or in the absence of IGF-I or GoH3 or rtIgG. Cell adhesion was analyzed by crystal violet staining. Data are expressed as mean ± SEM. (G) Surface expression of endogenous or exogenous β 1A or β 1C integrin or endogenous α 6 integrin was analyzed in PC3 cells by FACS ® using Ab to human β 1 , TS2/16, chicken β 1 , W1B10, α 6 , GoH3; or, as negative controls, mIgG, rtIgG, or 12CA5.
Figure Legend Snippet: β 1C enhances IGF-I– and II–mediated PC3 cell adhesion to LN-1. (A–D) β 1A - and β 1C -PC3 clones were cultured in the presence or in the absence of tet for 48 h. Cells were detached and plated on BSA or LN-1 or FN at 37°C for 2 h in the presence of IGF-II (A and B), of IGF-I (C and D), or of Ab to LN-1 (D) or ni-rIgG (D). In A, the differences in cell adhesion to LN-1 in the presence or in the absence of tet are statistically significant (*P ≤ 0.03). In C, the differences in cell adhesion to LN-1 between β 1A and β 1C expressing cells are statistically significant (*P ≤ 0.024). The experiments were repeated at least three times with similar results using two clones each of β 1A - and β 1C -PC3 cells. (E and F) β 1A - and β 1C -PC3 clones were cultured in the presence or in the absence of tet for 48 h. Cells were serum starved for 12 h, detached, and plated on LN-1 (E), FN or BSA (F) in serum-free medium at 37°C for 2 h in the presence or in the absence of IGF-I or GoH3 or rtIgG. Cell adhesion was analyzed by crystal violet staining. Data are expressed as mean ± SEM. (G) Surface expression of endogenous or exogenous β 1A or β 1C integrin or endogenous α 6 integrin was analyzed in PC3 cells by FACS ® using Ab to human β 1 , TS2/16, chicken β 1 , W1B10, α 6 , GoH3; or, as negative controls, mIgG, rtIgG, or 12CA5.

Techniques Used: Clone Assay, Cell Culture, Expressing, Staining, FACS

IGF stimulates adhesion to LN-1 of β 1C -expressing cells. β 1C -CHO (clones 16.3 and 16.4) and β 1A -CHO (clones 10.18 and 10.2) clones were cultured in the absence (A) or in the presence (B) of tet for 48 h. Cells were labeled using 51 Cr-sodium chromate for 1 h at 37°C. 51 Cr-labeled cells were incubated in the presence or in the absence of purified rabbit Ab to IGF-II or ni-rIgG for 1 h on ice. Cells were plated on BSA, LN-1, or FN at 37°C for 2 h in the presence or in the absence of IGF-II. Attached cells were washed, lysed, and the amount of 51 Cr associated with the attached cells was measured by liquid scintillation counting. Data are expressed as mean ± SEM. (C) Representative β 1C -CHO and β 1A -CHO clones were cultured in the absence or in the presence of tet for 48 h and analyzed by FACS ® using a β 1 integrin Ab TS2/16 specific to human β 1 or 7E2 specific to hamster β 1 or, as negative control, 12CA5.
Figure Legend Snippet: IGF stimulates adhesion to LN-1 of β 1C -expressing cells. β 1C -CHO (clones 16.3 and 16.4) and β 1A -CHO (clones 10.18 and 10.2) clones were cultured in the absence (A) or in the presence (B) of tet for 48 h. Cells were labeled using 51 Cr-sodium chromate for 1 h at 37°C. 51 Cr-labeled cells were incubated in the presence or in the absence of purified rabbit Ab to IGF-II or ni-rIgG for 1 h on ice. Cells were plated on BSA, LN-1, or FN at 37°C for 2 h in the presence or in the absence of IGF-II. Attached cells were washed, lysed, and the amount of 51 Cr associated with the attached cells was measured by liquid scintillation counting. Data are expressed as mean ± SEM. (C) Representative β 1C -CHO and β 1A -CHO clones were cultured in the absence or in the presence of tet for 48 h and analyzed by FACS ® using a β 1 integrin Ab TS2/16 specific to human β 1 or 7E2 specific to hamster β 1 or, as negative control, 12CA5.

Techniques Used: Expressing, Clone Assay, Cell Culture, Labeling, Incubation, Purification, FACS, Negative Control

9) Product Images from "Versatile ultrathin nanoporous silicon nitride membranes"

Article Title: Versatile ultrathin nanoporous silicon nitride membranes

Journal:

doi: 10.1073/pnas.0911450106

Permeation of Alexa Fluor, BSA, and IgG proteins through ITE SiN membranes. Fluorescent images were taken 4 min after introduction of the feed solution on top of the membrane. ( A ) Fluorescence images due to permeation of the Alexa dye ( Left ), BSA ( Center
Figure Legend Snippet: Permeation of Alexa Fluor, BSA, and IgG proteins through ITE SiN membranes. Fluorescent images were taken 4 min after introduction of the feed solution on top of the membrane. ( A ) Fluorescence images due to permeation of the Alexa dye ( Left ), BSA ( Center

Techniques Used: Fluorescence

10) Product Images from "Two pKM101-Encoded Proteins, the Pilus-Tip Protein TraC and Pep, Assemble on the Escherichia coli Cell Surface as Adhesins Required for Efficient Conjugative DNA Transfer"

Article Title: Two pKM101-Encoded Proteins, the Pilus-Tip Protein TraC and Pep, Assemble on the Escherichia coli Cell Surface as Adhesins Required for Efficient Conjugative DNA Transfer

Journal: Molecular microbiology

doi: 10.1111/mmi.14141

TraC and Pep localize on the E. coli cell surface. A) Upper: TraC ST is surface-displayed in the absence of the Tra pKM101 T4SS as determined with a dot blot assay. Strains: MC4100 producing the indicated proteins from plasmids in parantheses: TraC ST (pCGR83), ST TraC (pCGR66), MBP ST (pCGR84), TraF ST (pCGR48), untagged (WT) TraC (pCGR35). Whole cells were spotted onto nitrocellulose and probed for detection of Strep-tagged proteins (with α-Strep), surface-exposed RcsF lipoprotein (α-RcsF), or the periplasmic POTRA domains of BamA (α-BamA POTRA). Lower: Lysates from cells deposited onto the nitrocellulose were analyzed for steady-state accumulation of Strep-tagged TraC, MBP, and TraF by SDS-PAGE and immunostaining with α-Strep antibodies. Left: Molecular size markers (kDa). Blots were also developed with antibodies to the RNAP β subunit as a loading control. B) TraC.V144C ST reacts with BSA-Mal in the absence of the Tra pKM101 T4SS. Strains: MC4100 producing TraC ST (pDA107), TraC.Q46C ST (pDA110), or TraC.V144C ST (pDA112). Intact cells were treated (+) or not treated (−) with BSA-Mal, the reactions were quenched, and cell lysates were analyzed for formation of TraC ST /BSA-Mal complexes by SDS-PAGE and immunostaining with α-FLAG antibodies. Numbers at left: molecular size markers (kDa). Positions of higher-order TraC ST /BSA-Mal complexes and monomeric TraC ST are indicated. C) Identification of Pep FL in the extracellular fraction obtained by shearing of cells. Strains: MC4100 cells producing Pep FL (from pDA103), native Pep (pDA108), or MBP FL (pDA106). Whole cell extracts (left) or extracellular material recovered by shearing of cells (right) was analyzed for the presence of Pep FL , and periplasmic MBP FL as a control, by immunostaining with α-FLAG antibodies. A nonspecific crossreactive species detected in extracellular fractions from all strains served as a loading control. Immunoblots were also stained with antibodies reactive to the periplasmic POTRA domains of BamA and the cytosolic β subunit of RNA polymerase (RNAP). D) Pep is displayed on the cell surface in the dot blot assay. Strains: MC4100 producing Pep FL , Pep, or MBP FL from the above-listed plasmids, or Pep bearing FLAG tags immediately following the residue indicated from plasmids pDA113, 117, 115, 119, 116, 120, or 121. Whole cells were spotted onto nitrocellulose and probed with α-FLAG for detection of FLAG-tagged proteins, α-RcsF for surface-exposed RcsF lipoprotein, or α-BamA POTRA for periplasmic POTRA domains of BamA. Lysates from cells deposited onto the nitrocellulose were analyzed for Pep FL or MBP FL protein abundance by SDS-PAGE and immunostaining. Left: Molecular size markers (kDa). Blots were probed with α-RNAP for detection of cytosolic β-subunit of RNA polymerase as a loading control. E) PepC105 FL crosslinks with membrane-impermeable BSA-Mal. Strains: MC4100 producing Pep FL (pDA103) or Pep.C105 FL (pMB005). Intact cells were treated (+) or not treated (−) with BSA-Mal, the reactions were quenched, and cell lysates were analyzed for formation of Pep FL /BSA-Mal complexes by SDS-PAGE and immunostaining with α-FLAG antibodies. Right: Positions of higher-order Pep FL /BSA-Mal complexes and monomeric Pep FL are indicated. Left: Molecular size markers (kDa).
Figure Legend Snippet: TraC and Pep localize on the E. coli cell surface. A) Upper: TraC ST is surface-displayed in the absence of the Tra pKM101 T4SS as determined with a dot blot assay. Strains: MC4100 producing the indicated proteins from plasmids in parantheses: TraC ST (pCGR83), ST TraC (pCGR66), MBP ST (pCGR84), TraF ST (pCGR48), untagged (WT) TraC (pCGR35). Whole cells were spotted onto nitrocellulose and probed for detection of Strep-tagged proteins (with α-Strep), surface-exposed RcsF lipoprotein (α-RcsF), or the periplasmic POTRA domains of BamA (α-BamA POTRA). Lower: Lysates from cells deposited onto the nitrocellulose were analyzed for steady-state accumulation of Strep-tagged TraC, MBP, and TraF by SDS-PAGE and immunostaining with α-Strep antibodies. Left: Molecular size markers (kDa). Blots were also developed with antibodies to the RNAP β subunit as a loading control. B) TraC.V144C ST reacts with BSA-Mal in the absence of the Tra pKM101 T4SS. Strains: MC4100 producing TraC ST (pDA107), TraC.Q46C ST (pDA110), or TraC.V144C ST (pDA112). Intact cells were treated (+) or not treated (−) with BSA-Mal, the reactions were quenched, and cell lysates were analyzed for formation of TraC ST /BSA-Mal complexes by SDS-PAGE and immunostaining with α-FLAG antibodies. Numbers at left: molecular size markers (kDa). Positions of higher-order TraC ST /BSA-Mal complexes and monomeric TraC ST are indicated. C) Identification of Pep FL in the extracellular fraction obtained by shearing of cells. Strains: MC4100 cells producing Pep FL (from pDA103), native Pep (pDA108), or MBP FL (pDA106). Whole cell extracts (left) or extracellular material recovered by shearing of cells (right) was analyzed for the presence of Pep FL , and periplasmic MBP FL as a control, by immunostaining with α-FLAG antibodies. A nonspecific crossreactive species detected in extracellular fractions from all strains served as a loading control. Immunoblots were also stained with antibodies reactive to the periplasmic POTRA domains of BamA and the cytosolic β subunit of RNA polymerase (RNAP). D) Pep is displayed on the cell surface in the dot blot assay. Strains: MC4100 producing Pep FL , Pep, or MBP FL from the above-listed plasmids, or Pep bearing FLAG tags immediately following the residue indicated from plasmids pDA113, 117, 115, 119, 116, 120, or 121. Whole cells were spotted onto nitrocellulose and probed with α-FLAG for detection of FLAG-tagged proteins, α-RcsF for surface-exposed RcsF lipoprotein, or α-BamA POTRA for periplasmic POTRA domains of BamA. Lysates from cells deposited onto the nitrocellulose were analyzed for Pep FL or MBP FL protein abundance by SDS-PAGE and immunostaining. Left: Molecular size markers (kDa). Blots were probed with α-RNAP for detection of cytosolic β-subunit of RNA polymerase as a loading control. E) PepC105 FL crosslinks with membrane-impermeable BSA-Mal. Strains: MC4100 producing Pep FL (pDA103) or Pep.C105 FL (pMB005). Intact cells were treated (+) or not treated (−) with BSA-Mal, the reactions were quenched, and cell lysates were analyzed for formation of Pep FL /BSA-Mal complexes by SDS-PAGE and immunostaining with α-FLAG antibodies. Right: Positions of higher-order Pep FL /BSA-Mal complexes and monomeric Pep FL are indicated. Left: Molecular size markers (kDa).

Techniques Used: Dot Blot, SDS Page, Immunostaining, Western Blot, Staining

11) Product Images from "Members of the Pmp protein family of Chlamydia pneumoniae mediate adhesion to human cells via short repetitive peptide motifs"

Article Title: Members of the Pmp protein family of Chlamydia pneumoniae mediate adhesion to human cells via short repetitive peptide motifs

Journal: Molecular Microbiology

doi: 10.1111/j.1365-2958.2010.07386.x

Pmp6, Pmp20 and Pmp21 carry multiple repeats of both tetrapeptide motifs, adhere to HEp-2 cells and are important for infection.A. Schematic representation of the Pmp6, Pmp20 and Pmp21 proteins from C. pneumoniae , together with the protein domains analysed here. The position of each of the tetrapeptide motifs GGA(I, L, V) and FxxN is marked and the total number of each motif type found is given on the right.B. Adhesion of Pmp6- or Pmp20-coated fluorescent latex beads to HEp-2 cells. Green fluorescent beads (1 × 10 7 ) were coated with (200 µg ml −1 ) BSA, recombinant Inv 497 (rInv 497 ), rPmp6-1, rPmp20-1 or rPmp21-E, and incubated with 1 × 10 6 HEp-2 cells. Mean fluorescence of HEp-2 cells was determined by flow cytometric analysis. (1 × 10 6 HEp-2 cells per sample). Data are based on four independent experiments for each protein.C. Adhesion of viable, CFSE-stained C. pneumoniae EBs to human HEp-2 cells is reduced in the presence of rPmp6, rPmp20 or rPmp21. The fluorescence intensity associated with attachment of EBs to HEp-2 cells in the presence of PBS served as the control and was set to 100%. Data are based on two independent experiments, each performed in triplicate ( n = 6). Error bars indicate standard deviations.
Figure Legend Snippet: Pmp6, Pmp20 and Pmp21 carry multiple repeats of both tetrapeptide motifs, adhere to HEp-2 cells and are important for infection.A. Schematic representation of the Pmp6, Pmp20 and Pmp21 proteins from C. pneumoniae , together with the protein domains analysed here. The position of each of the tetrapeptide motifs GGA(I, L, V) and FxxN is marked and the total number of each motif type found is given on the right.B. Adhesion of Pmp6- or Pmp20-coated fluorescent latex beads to HEp-2 cells. Green fluorescent beads (1 × 10 7 ) were coated with (200 µg ml −1 ) BSA, recombinant Inv 497 (rInv 497 ), rPmp6-1, rPmp20-1 or rPmp21-E, and incubated with 1 × 10 6 HEp-2 cells. Mean fluorescence of HEp-2 cells was determined by flow cytometric analysis. (1 × 10 6 HEp-2 cells per sample). Data are based on four independent experiments for each protein.C. Adhesion of viable, CFSE-stained C. pneumoniae EBs to human HEp-2 cells is reduced in the presence of rPmp6, rPmp20 or rPmp21. The fluorescence intensity associated with attachment of EBs to HEp-2 cells in the presence of PBS served as the control and was set to 100%. Data are based on two independent experiments, each performed in triplicate ( n = 6). Error bars indicate standard deviations.

Techniques Used: Infection, Recombinant, Incubation, Fluorescence, Flow Cytometry, Staining

Recombinant Pmp21 inhibits infection by C. pneumoniae . Infection of HEp-2 cells by C. pneumoniae is inhibited by prior addition of anti-Pmp21, recombinant Pmp21 or a synthetic peptide derived from Pmp21.A. Purified EBs were incubated with PBS, pre-immune serum (diluted 1:1 with PBS) or serially diluted anti-Pmp21-E before being used for infection of HEp-2 cells. The number of inclusions formed was determined by microscopy 48 h p.i. The data in (A), (B) and (D) are derived from four independent experiments for each condition, each involving observation of 20 microscope fields.B. HEp-2 cells (1 × 10 6 ) were incubated with PBS, 200 µg ml −1 BSA, or 100 µg ml −1 or 200 µg ml −1 recombinant Pmp21-PD or one of the other Pmp21 variants as indicated prior to incubation with purified C. pneumoniae EBs. Cells were fixed 48 h p.i. and the number of inclusions determined as explained above. PBS control = 100%.C. Adhesion of viable, CFSE-stained C. pneumoniae EBs to human HEp-2 cells was detected by flow cytometry. Attachment of EBs in the presence of PBS or 200 µg ml −1 BSA served as the control, and the fluorescence intensity associated with HEp-2 cells pre-incubated in PBS was set to 100%. Binding of EBs in the presence of 500 U ml −1 heparin or increasing amounts of recombinant Pmp21-PD were analysed accordingly. n = 2 wells; number of experiments = 6. Error bars indicate standard deviations.D. A Pmp21-derived peptide attenuates C. pneumoniae infection. Prior to incubation with purified C. pneumoniae EBs, HEp-2 cells (1 × 10 6 ) were incubated without or with increasing amounts (1.7–54.4 µg ml −1 ) of a 32-amino-acid peptide derived from Pmp21 (residues 745–776) (peptide) or a scrambled version of the sequence with the same amino acid composition (scrambled peptide). Cells were fixed 48 h p.i. and the number of inclusions determined by microscopy as detailed above. PBS control = 100%.
Figure Legend Snippet: Recombinant Pmp21 inhibits infection by C. pneumoniae . Infection of HEp-2 cells by C. pneumoniae is inhibited by prior addition of anti-Pmp21, recombinant Pmp21 or a synthetic peptide derived from Pmp21.A. Purified EBs were incubated with PBS, pre-immune serum (diluted 1:1 with PBS) or serially diluted anti-Pmp21-E before being used for infection of HEp-2 cells. The number of inclusions formed was determined by microscopy 48 h p.i. The data in (A), (B) and (D) are derived from four independent experiments for each condition, each involving observation of 20 microscope fields.B. HEp-2 cells (1 × 10 6 ) were incubated with PBS, 200 µg ml −1 BSA, or 100 µg ml −1 or 200 µg ml −1 recombinant Pmp21-PD or one of the other Pmp21 variants as indicated prior to incubation with purified C. pneumoniae EBs. Cells were fixed 48 h p.i. and the number of inclusions determined as explained above. PBS control = 100%.C. Adhesion of viable, CFSE-stained C. pneumoniae EBs to human HEp-2 cells was detected by flow cytometry. Attachment of EBs in the presence of PBS or 200 µg ml −1 BSA served as the control, and the fluorescence intensity associated with HEp-2 cells pre-incubated in PBS was set to 100%. Binding of EBs in the presence of 500 U ml −1 heparin or increasing amounts of recombinant Pmp21-PD were analysed accordingly. n = 2 wells; number of experiments = 6. Error bars indicate standard deviations.D. A Pmp21-derived peptide attenuates C. pneumoniae infection. Prior to incubation with purified C. pneumoniae EBs, HEp-2 cells (1 × 10 6 ) were incubated without or with increasing amounts (1.7–54.4 µg ml −1 ) of a 32-amino-acid peptide derived from Pmp21 (residues 745–776) (peptide) or a scrambled version of the sequence with the same amino acid composition (scrambled peptide). Cells were fixed 48 h p.i. and the number of inclusions determined by microscopy as detailed above. PBS control = 100%.

Techniques Used: Recombinant, Infection, Derivative Assay, Purification, Incubation, Microscopy, Staining, Flow Cytometry, Cytometry, Fluorescence, Binding Assay, Sequencing

12) Product Images from "CaMKII Binding to GluN2B Is Differentially Affected by Macromolecular Crowding Reagents"

Article Title: CaMKII Binding to GluN2B Is Differentially Affected by Macromolecular Crowding Reagents

Journal: PLoS ONE

doi: 10.1371/journal.pone.0096522

Ca 2+ /CaM stimulation is still required for CaMKII to GluN2B binding. A , CaMKIIα (40 nM subunits) was incubated with immuno-immobilized GST-GluN2B-C as in Figure 1A , but in the presence or absence of Ca 2+ /CaM (1 mM/1 µM), ADP (100 µM), or BSA (100 mg/ml). Bound CaMKII was eluted and detected by Western-analysis. BSA and ADP alone were insufficient to induce CaMKII to GluN2B binding. GST-GluN2B detection is shown as a loading control. Representative images are from the same experiment and Western-blots. B , CaMKIIα (40 nM subunits) was incubated with immuno-immobilized GST-GluN2B-C as in panel A. Different crowding agents, BSA, dextran-10 (DEX10) and dextran-70 (DEX70) (all at 100 mg/ml) in the presence of ADP (100 µM) were not sufficient to induce detectable CaMKII binding to GluN2B without CaMKII stimulation.
Figure Legend Snippet: Ca 2+ /CaM stimulation is still required for CaMKII to GluN2B binding. A , CaMKIIα (40 nM subunits) was incubated with immuno-immobilized GST-GluN2B-C as in Figure 1A , but in the presence or absence of Ca 2+ /CaM (1 mM/1 µM), ADP (100 µM), or BSA (100 mg/ml). Bound CaMKII was eluted and detected by Western-analysis. BSA and ADP alone were insufficient to induce CaMKII to GluN2B binding. GST-GluN2B detection is shown as a loading control. Representative images are from the same experiment and Western-blots. B , CaMKIIα (40 nM subunits) was incubated with immuno-immobilized GST-GluN2B-C as in panel A. Different crowding agents, BSA, dextran-10 (DEX10) and dextran-70 (DEX70) (all at 100 mg/ml) in the presence of ADP (100 µM) were not sufficient to induce detectable CaMKII binding to GluN2B without CaMKII stimulation.

Techniques Used: Chick Chorioallantoic Membrane Assay, Binding Assay, Incubation, Western Blot

Lysozyme but not BSA binds to the GST immobilization scaffold. 100 mg/ml Lysozyme (6.8 mM) or BSA (1.5 mM) were tested for binding to immuno-immobilized GST-GluN2B-C under the same conditions as in Figure 1A . Samples were eluted and subjected to SDS-PAGE, after which the gel was fixed and stained for total protein (silver stain). Binding was detected for lysozyme but not BSA. Lysozyme binding was detected also to immobilized GST (without GluN2B fusion) and to empty wells, indicating non-specific binding to the immobilization scaffold.
Figure Legend Snippet: Lysozyme but not BSA binds to the GST immobilization scaffold. 100 mg/ml Lysozyme (6.8 mM) or BSA (1.5 mM) were tested for binding to immuno-immobilized GST-GluN2B-C under the same conditions as in Figure 1A . Samples were eluted and subjected to SDS-PAGE, after which the gel was fixed and stained for total protein (silver stain). Binding was detected for lysozyme but not BSA. Lysozyme binding was detected also to immobilized GST (without GluN2B fusion) and to empty wells, indicating non-specific binding to the immobilization scaffold.

Techniques Used: Binding Assay, SDS Page, Staining, Silver Staining

Differential effects of BSA and lysozyme molecular crowding agents in CaMKII to GluN2B binding. A , CaMKIIα (40 nM subunits) binding to GST-GluN2B-C that was immuno-immobilized on anti-GST coated microtiter wells was induced by Ca 2+ /CaM (1 mM/1 µM) in the presence of ADP (100 µM) for 15 min at room temperature. Bound CaMKII was eluted and detected by Western-analysis, and quantified by normalized immuno-detection values (IDV). Macromolecular crowding with lysozyme (100 mg/ml) decreased CaMKII binding to GluN2B, while BSA (100 mg/ml) instead increased binding. n = 4; *: p
Figure Legend Snippet: Differential effects of BSA and lysozyme molecular crowding agents in CaMKII to GluN2B binding. A , CaMKIIα (40 nM subunits) binding to GST-GluN2B-C that was immuno-immobilized on anti-GST coated microtiter wells was induced by Ca 2+ /CaM (1 mM/1 µM) in the presence of ADP (100 µM) for 15 min at room temperature. Bound CaMKII was eluted and detected by Western-analysis, and quantified by normalized immuno-detection values (IDV). Macromolecular crowding with lysozyme (100 mg/ml) decreased CaMKII binding to GluN2B, while BSA (100 mg/ml) instead increased binding. n = 4; *: p

Techniques Used: Binding Assay, Chick Chorioallantoic Membrane Assay, Western Blot

Lysozyme but not BSA binds Ca 2+ /CaM. A , The chicken lysozyme amino acid sequence with predicted CaM-binding sites marked. Prediction utilized the “Calmodulin Target Database” ( http://calcium.uhnres.utoronto.ca/ctdb/ctdb/home.html ). Numbers from 1 to 9 indicate increasing likelihood of predicted CaM binding. B , Lysozyme and BSA (0, 1, 2, 4, and 8 µg) were subjected to SDS-PAGE and transferred to a PVDF membrane. The membrane was first stained for total protein (ponceau, bottom panel), then incubated with biotin-labeled CaM in presence of CaCl 2 . Bound CaM was detected by chemi-luminescence (top panel). Ca 2+ /CaM bound Lysozyme, but not BSA.
Figure Legend Snippet: Lysozyme but not BSA binds Ca 2+ /CaM. A , The chicken lysozyme amino acid sequence with predicted CaM-binding sites marked. Prediction utilized the “Calmodulin Target Database” ( http://calcium.uhnres.utoronto.ca/ctdb/ctdb/home.html ). Numbers from 1 to 9 indicate increasing likelihood of predicted CaM binding. B , Lysozyme and BSA (0, 1, 2, 4, and 8 µg) were subjected to SDS-PAGE and transferred to a PVDF membrane. The membrane was first stained for total protein (ponceau, bottom panel), then incubated with biotin-labeled CaM in presence of CaCl 2 . Bound CaM was detected by chemi-luminescence (top panel). Ca 2+ /CaM bound Lysozyme, but not BSA.

Techniques Used: Chick Chorioallantoic Membrane Assay, Sequencing, Binding Assay, SDS Page, Staining, Incubation, Labeling

13) Product Images from "Dectin-2-Targeted Antifungal Liposomes Exhibit Enhanced Efficacy"

Article Title: Dectin-2-Targeted Antifungal Liposomes Exhibit Enhanced Efficacy

Journal: mSphere

doi: 10.1128/mSphere.00715-19

sDectin-2-coated amphotericin B-loaded liposome inhibition and killing of C. albicans , C. neoformans , and A. fumigatus . (A) C. albicans with DEC2-AmB-LLs. Cells in the pseudohyphal and early hyphal stage grown in RPMI medium plus 0.5% BSA on 96-well polystyrene microtiter plates. Cells were treated for 30 min with liposomes delivering 1.0., 0.5, 0.25, and 0.12 μM AmB to the medium as indicated, washed twice with medium, grown for an additional 16 h, and then assayed for metabolic activity using CellTiter-Blue (CTB) reagent. (B) C. neoformans with DEC2-AmB-LLs. C. neoformans cells were grown in liquid YPD medium plus 0.5% BSA with vigorous shaking for 2 h and treated for 4 h or overnight with liposomes delivering 0.4, 0.2, or 0.1 μM AmB to the medium as indicated. Cells were diluted and plated on YPD medium, and CFU were counted from multiple plates. (C) A. fumigatus with DEC2-AmB-LLs. Conidia were germinated for 9 h in VMM plus glucose plus 0.5% BSA in 96-well polystyrene microtiter plates, treated for 2 h with liposomes delivering 0.5 and 0.25 μM AmB to the medium as indicated, washed twice with medium, grown overnight, and then assayed for metabolic activity using CTB reagent in RPMI lacking phenol red indicator plus 0.5% BSA. The control wells were overgrown with hyphae protruding out the medium. (D) A. fumigatus with DEC1-AmB-LLs. Assay conditions were similar to the assay in panel C, except liposomes were first diluted into LDB1 buffer (PBS plus 0.5%BSA plus 1 mM BME) prior to dilution into growth medium. For CTB assays in panels A, C, and D, the fluorescence background from medium incubated with CTB reagent was subtracted. Standard errors are shown for all values, and fold differences and P values were estimated comparing the performance of AmB-LLs to DEC2-AmB-LLs. Two or more biological replicates gave similar results.
Figure Legend Snippet: sDectin-2-coated amphotericin B-loaded liposome inhibition and killing of C. albicans , C. neoformans , and A. fumigatus . (A) C. albicans with DEC2-AmB-LLs. Cells in the pseudohyphal and early hyphal stage grown in RPMI medium plus 0.5% BSA on 96-well polystyrene microtiter plates. Cells were treated for 30 min with liposomes delivering 1.0., 0.5, 0.25, and 0.12 μM AmB to the medium as indicated, washed twice with medium, grown for an additional 16 h, and then assayed for metabolic activity using CellTiter-Blue (CTB) reagent. (B) C. neoformans with DEC2-AmB-LLs. C. neoformans cells were grown in liquid YPD medium plus 0.5% BSA with vigorous shaking for 2 h and treated for 4 h or overnight with liposomes delivering 0.4, 0.2, or 0.1 μM AmB to the medium as indicated. Cells were diluted and plated on YPD medium, and CFU were counted from multiple plates. (C) A. fumigatus with DEC2-AmB-LLs. Conidia were germinated for 9 h in VMM plus glucose plus 0.5% BSA in 96-well polystyrene microtiter plates, treated for 2 h with liposomes delivering 0.5 and 0.25 μM AmB to the medium as indicated, washed twice with medium, grown overnight, and then assayed for metabolic activity using CTB reagent in RPMI lacking phenol red indicator plus 0.5% BSA. The control wells were overgrown with hyphae protruding out the medium. (D) A. fumigatus with DEC1-AmB-LLs. Assay conditions were similar to the assay in panel C, except liposomes were first diluted into LDB1 buffer (PBS plus 0.5%BSA plus 1 mM BME) prior to dilution into growth medium. For CTB assays in panels A, C, and D, the fluorescence background from medium incubated with CTB reagent was subtracted. Standard errors are shown for all values, and fold differences and P values were estimated comparing the performance of AmB-LLs to DEC2-AmB-LLs. Two or more biological replicates gave similar results.

Techniques Used: Inhibition, Activity Assay, CtB Assay, Fluorescence, Incubation

14) Product Images from "GSK3β mediates pancreatic cancer cell invasion in vitro via the CXCR4/MMP-2 Pathway"

Article Title: GSK3β mediates pancreatic cancer cell invasion in vitro via the CXCR4/MMP-2 Pathway

Journal: Cancer Cell International

doi: 10.1186/s12935-015-0216-y

GSK3β promotes PANC1 human pancreatic cancer cells invasion. Transwell invasion assay was employed to determine the effect of GSK3β on cell invasion. Stable cell clones of PANC1 cells overexpression or suppression of GSK3β were selected. 200 μl cell suspension maitained in serum-free DMEM/0.1 % BSA were added to the matrigel-coated upper compartment, the DMED/10%FBS medium was added to the lower compartment, and the plates were incubated for 48 h at 37 °C. a After incubation for 48 h, the upper surface of the filter was scrubbed free of cells, the filter was fixed and stained and the lower surface was photographed (200× original magnification). b Quantification of A. Results are expressed as the mean ± SD; * p
Figure Legend Snippet: GSK3β promotes PANC1 human pancreatic cancer cells invasion. Transwell invasion assay was employed to determine the effect of GSK3β on cell invasion. Stable cell clones of PANC1 cells overexpression or suppression of GSK3β were selected. 200 μl cell suspension maitained in serum-free DMEM/0.1 % BSA were added to the matrigel-coated upper compartment, the DMED/10%FBS medium was added to the lower compartment, and the plates were incubated for 48 h at 37 °C. a After incubation for 48 h, the upper surface of the filter was scrubbed free of cells, the filter was fixed and stained and the lower surface was photographed (200× original magnification). b Quantification of A. Results are expressed as the mean ± SD; * p

Techniques Used: Transwell Invasion Assay, Stable Transfection, Clone Assay, Over Expression, Incubation, Staining

15) Product Images from "Transthyretin Maintains Muscle Homeostasis through the Novel Shuttle Pathway of Thyroid Hormones during Myoblast Differentiation"

Article Title: Transthyretin Maintains Muscle Homeostasis through the Novel Shuttle Pathway of Thyroid Hormones during Myoblast Differentiation

Journal: Cells

doi: 10.3390/cells8121565

Endocytosis of TTR protein and TTR overexpression effects. ( A ) TTR protein or BSA were labeled with fluorescence and cells were cultured with serum-free media supplemented with labeled TTR protein or BSA for 1 day. Detection of labeled TTR protein and BSA in cells (Red: TTR, Blue: Nucleus). ( B ) TTR overexpression was performed by transfecting with TTR ORF plasmid followed by incubation with 10% FBS for two days. Cell viability was analyzed by MTT assay. ( C ) TTR overexpressing cells were incubated with serum-free media for two days. Myotube formation and fusion index were observed by Giemsa staining, TTR mRNA level by real-time RT-PCR, and protein expression by Western blot and immunocytochemistry. Control or TTR-overexpressing cells were incubated with serum-free media supplemented with T 4 for two days. T 4 or T 3 concentration was measured by ELISA. Means ± SD ( n = 3). * p ≤ 0.05, ** p ≤ 0.001, *** p ≤ 0.0001.
Figure Legend Snippet: Endocytosis of TTR protein and TTR overexpression effects. ( A ) TTR protein or BSA were labeled with fluorescence and cells were cultured with serum-free media supplemented with labeled TTR protein or BSA for 1 day. Detection of labeled TTR protein and BSA in cells (Red: TTR, Blue: Nucleus). ( B ) TTR overexpression was performed by transfecting with TTR ORF plasmid followed by incubation with 10% FBS for two days. Cell viability was analyzed by MTT assay. ( C ) TTR overexpressing cells were incubated with serum-free media for two days. Myotube formation and fusion index were observed by Giemsa staining, TTR mRNA level by real-time RT-PCR, and protein expression by Western blot and immunocytochemistry. Control or TTR-overexpressing cells were incubated with serum-free media supplemented with T 4 for two days. T 4 or T 3 concentration was measured by ELISA. Means ± SD ( n = 3). * p ≤ 0.05, ** p ≤ 0.001, *** p ≤ 0.0001.

Techniques Used: Over Expression, Labeling, Fluorescence, Cell Culture, Plasmid Preparation, Incubation, MTT Assay, Staining, Quantitative RT-PCR, Expressing, Western Blot, Immunocytochemistry, Concentration Assay, Enzyme-linked Immunosorbent Assay

Myoblast differentiation following BSA treatment. Cells were cultured in serum-free media supplemented with T 4 or T 4 + BSA for two days ( A – E ). ( A ) Myotube formation and fusion index were observed by Giemsa staining. mRNA level was observed by real-time RT-PCR and protein expressions by Western blot and immunocytochemistry. ( B ) TTR mRNA in exosomes of cultured media using RT-PCR and protein level in cultured media by Western blot. ( C ) T 4 or T 3 concentration in cultured media or cells was observed by ELISA. ( D , E ) Cells were cultured with serum-free media supplemented with T 4 , T 4 + BSA, T 4 + TTR or T 4 + BSA + TTR for two days. T 4 or T 3 concentration in T 4 + BSA or T 4 + BSA + TTR treated cells. TTR mRNA in exosomes of cultured media (in T 4 + BSA or T 4 + BSA + TTR treated cells) using RT-PCR. ( F ) Cells were cultured in serum-free media supplemented with T 4 or T 4 + BSA or T 4 + TTR for two days and exosomes were isolated from each cultured medium. T 3 concentration in exosomes. Means ± SD ( n = 3). * p ≤ 0.05, ** p ≤ 0.001, *** p ≤ 0.0001.
Figure Legend Snippet: Myoblast differentiation following BSA treatment. Cells were cultured in serum-free media supplemented with T 4 or T 4 + BSA for two days ( A – E ). ( A ) Myotube formation and fusion index were observed by Giemsa staining. mRNA level was observed by real-time RT-PCR and protein expressions by Western blot and immunocytochemistry. ( B ) TTR mRNA in exosomes of cultured media using RT-PCR and protein level in cultured media by Western blot. ( C ) T 4 or T 3 concentration in cultured media or cells was observed by ELISA. ( D , E ) Cells were cultured with serum-free media supplemented with T 4 , T 4 + BSA, T 4 + TTR or T 4 + BSA + TTR for two days. T 4 or T 3 concentration in T 4 + BSA or T 4 + BSA + TTR treated cells. TTR mRNA in exosomes of cultured media (in T 4 + BSA or T 4 + BSA + TTR treated cells) using RT-PCR. ( F ) Cells were cultured in serum-free media supplemented with T 4 or T 4 + BSA or T 4 + TTR for two days and exosomes were isolated from each cultured medium. T 3 concentration in exosomes. Means ± SD ( n = 3). * p ≤ 0.05, ** p ≤ 0.001, *** p ≤ 0.0001.

Techniques Used: Cell Culture, Staining, Quantitative RT-PCR, Western Blot, Immunocytochemistry, Reverse Transcription Polymerase Chain Reaction, Concentration Assay, Enzyme-linked Immunosorbent Assay, Isolation

16) Product Images from "CaMKII Binding to GluN2B Is Differentially Affected by Macromolecular Crowding Reagents"

Article Title: CaMKII Binding to GluN2B Is Differentially Affected by Macromolecular Crowding Reagents

Journal: PLoS ONE

doi: 10.1371/journal.pone.0096522

Ca 2+ /CaM stimulation is still required for CaMKII to GluN2B binding. A , CaMKIIα (40 nM subunits) was incubated with immuno-immobilized GST-GluN2B-C as in Figure 1A , but in the presence or absence of Ca 2+ /CaM (1 mM/1 µM), ADP (100 µM), or BSA (100 mg/ml). Bound CaMKII was eluted and detected by Western-analysis. BSA and ADP alone were insufficient to induce CaMKII to GluN2B binding. GST-GluN2B detection is shown as a loading control. Representative images are from the same experiment and Western-blots. B , CaMKIIα (40 nM subunits) was incubated with immuno-immobilized GST-GluN2B-C as in panel A. Different crowding agents, BSA, dextran-10 (DEX10) and dextran-70 (DEX70) (all at 100 mg/ml) in the presence of ADP (100 µM) were not sufficient to induce detectable CaMKII binding to GluN2B without CaMKII stimulation.
Figure Legend Snippet: Ca 2+ /CaM stimulation is still required for CaMKII to GluN2B binding. A , CaMKIIα (40 nM subunits) was incubated with immuno-immobilized GST-GluN2B-C as in Figure 1A , but in the presence or absence of Ca 2+ /CaM (1 mM/1 µM), ADP (100 µM), or BSA (100 mg/ml). Bound CaMKII was eluted and detected by Western-analysis. BSA and ADP alone were insufficient to induce CaMKII to GluN2B binding. GST-GluN2B detection is shown as a loading control. Representative images are from the same experiment and Western-blots. B , CaMKIIα (40 nM subunits) was incubated with immuno-immobilized GST-GluN2B-C as in panel A. Different crowding agents, BSA, dextran-10 (DEX10) and dextran-70 (DEX70) (all at 100 mg/ml) in the presence of ADP (100 µM) were not sufficient to induce detectable CaMKII binding to GluN2B without CaMKII stimulation.

Techniques Used: Chick Chorioallantoic Membrane Assay, Binding Assay, Incubation, Western Blot

Lysozyme but not BSA binds to the GST immobilization scaffold. 100 mg/ml Lysozyme (6.8 mM) or BSA (1.5 mM) were tested for binding to immuno-immobilized GST-GluN2B-C under the same conditions as in Figure 1A . Samples were eluted and subjected to SDS-PAGE, after which the gel was fixed and stained for total protein (silver stain). Binding was detected for lysozyme but not BSA. Lysozyme binding was detected also to immobilized GST (without GluN2B fusion) and to empty wells, indicating non-specific binding to the immobilization scaffold.
Figure Legend Snippet: Lysozyme but not BSA binds to the GST immobilization scaffold. 100 mg/ml Lysozyme (6.8 mM) or BSA (1.5 mM) were tested for binding to immuno-immobilized GST-GluN2B-C under the same conditions as in Figure 1A . Samples were eluted and subjected to SDS-PAGE, after which the gel was fixed and stained for total protein (silver stain). Binding was detected for lysozyme but not BSA. Lysozyme binding was detected also to immobilized GST (without GluN2B fusion) and to empty wells, indicating non-specific binding to the immobilization scaffold.

Techniques Used: Binding Assay, SDS Page, Staining, Silver Staining

Differential effects of BSA and lysozyme molecular crowding agents in CaMKII to GluN2B binding. A , CaMKIIα (40 nM subunits) binding to GST-GluN2B-C that was immuno-immobilized on anti-GST coated microtiter wells was induced by Ca 2+ /CaM (1 mM/1 µM) in the presence of ADP (100 µM) for 15 min at room temperature. Bound CaMKII was eluted and detected by Western-analysis, and quantified by normalized immuno-detection values (IDV). Macromolecular crowding with lysozyme (100 mg/ml) decreased CaMKII binding to GluN2B, while BSA (100 mg/ml) instead increased binding. n = 4; *: p
Figure Legend Snippet: Differential effects of BSA and lysozyme molecular crowding agents in CaMKII to GluN2B binding. A , CaMKIIα (40 nM subunits) binding to GST-GluN2B-C that was immuno-immobilized on anti-GST coated microtiter wells was induced by Ca 2+ /CaM (1 mM/1 µM) in the presence of ADP (100 µM) for 15 min at room temperature. Bound CaMKII was eluted and detected by Western-analysis, and quantified by normalized immuno-detection values (IDV). Macromolecular crowding with lysozyme (100 mg/ml) decreased CaMKII binding to GluN2B, while BSA (100 mg/ml) instead increased binding. n = 4; *: p

Techniques Used: Binding Assay, Chick Chorioallantoic Membrane Assay, Western Blot

Lysozyme but not BSA binds Ca 2+ /CaM. A , The chicken lysozyme amino acid sequence with predicted CaM-binding sites marked. Prediction utilized the “Calmodulin Target Database” ( http://calcium.uhnres.utoronto.ca/ctdb/ctdb/home.html ). Numbers from 1 to 9 indicate increasing likelihood of predicted CaM binding. B , Lysozyme and BSA (0, 1, 2, 4, and 8 µg) were subjected to SDS-PAGE and transferred to a PVDF membrane. The membrane was first stained for total protein (ponceau, bottom panel), then incubated with biotin-labeled CaM in presence of CaCl 2 . Bound CaM was detected by chemi-luminescence (top panel). Ca 2+ /CaM bound Lysozyme, but not BSA.
Figure Legend Snippet: Lysozyme but not BSA binds Ca 2+ /CaM. A , The chicken lysozyme amino acid sequence with predicted CaM-binding sites marked. Prediction utilized the “Calmodulin Target Database” ( http://calcium.uhnres.utoronto.ca/ctdb/ctdb/home.html ). Numbers from 1 to 9 indicate increasing likelihood of predicted CaM binding. B , Lysozyme and BSA (0, 1, 2, 4, and 8 µg) were subjected to SDS-PAGE and transferred to a PVDF membrane. The membrane was first stained for total protein (ponceau, bottom panel), then incubated with biotin-labeled CaM in presence of CaCl 2 . Bound CaM was detected by chemi-luminescence (top panel). Ca 2+ /CaM bound Lysozyme, but not BSA.

Techniques Used: Chick Chorioallantoic Membrane Assay, Sequencing, Binding Assay, SDS Page, Staining, Incubation, Labeling

17) Product Images from "Enterobactin-Specific Antibodies Induced by a Novel Enterobactin Conjugate Vaccine"

Article Title: Enterobactin-Specific Antibodies Induced by a Novel Enterobactin Conjugate Vaccine

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.00358-19

Ent conjugation with carrier proteins. (A) Schematic description of the conjugation of Ent to a carrier protein. (B) SDS-PAGE analysis of Ent conjugation with carrier protein KLH (left panel), BSA (middle panel), or CmeC (right panel).
Figure Legend Snippet: Ent conjugation with carrier proteins. (A) Schematic description of the conjugation of Ent to a carrier protein. (B) SDS-PAGE analysis of Ent conjugation with carrier protein KLH (left panel), BSA (middle panel), or CmeC (right panel).

Techniques Used: Conjugation Assay, SDS Page

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Article Snippet: .. Flow Cytometry Analyses After staining the THP-1 cells in an Invitrogen LIVE/DEAD Fixable Red Dead Cell Stain kit (#L23102, Thermo Fisher Scientific Inc., Waltham, MA, USA), viable cells were blocked using 10μg/mL human immunoglobulin G (IgG) diluted in 0.5% bovine serum albumin (BSA)/1mM, ethylenediaminetetraacetic acid (EDTA)/0.01%, sodium azide staining buffer on ice for 10 min, and then incubated with anti-human CD206 (MMR)-PE antibody (#12-2069-41, 1:50, eBioscience, Thermo Fisher Scientific Inc., Waltham, MA, USA) on ice for 1 h, before cell fixation in 2% paraformaldehyde (PFA) on ice for 15 min. Next, the cells were incubated with anti-human CD80 (B7-1) Fluorescein isothiocyanate (FITC) antibodies (#11-0809-41, 1:50, eBioscience, Thermo Fisher Scientific Inc., Waltham, MA, USA) in 0.5% saponin on ice for 45 min, washed with staining buffer, and then flow-cytometry data acquisition was performed using the BD FACSCanto II system (BD Biosciences, Thermo Fisher Scientific Inc. Waltham, MA, USA). .. Data obtained were analyzed using Flowjo software v. 9.6.2 (FlowJo, LLC, Tree Star, Inc., Ashland, OR, USA).

Article Title: Enzyme-responsive nanocomposites for wound infection prophylaxis in burn management: in vitro evaluation of their compatibility with healing processes
Article Snippet: .. Subsequently, secondary antibody (Alexa Fluor 488 and 594, Life Technologies, Carlsbad, CA, USA, 1:1,000 in 1% bovine serum albumin in PBS) was added and incubated for 60 minutes. .. After washing three times with 0.05% Tween 20 in PBS, nuclear counterstaining was performed by adding DAPI (Sigma-Aldrich) (1:10,000 dilution in PBS) for 5 minutes.

Article Title: SPINK1 is a prognosis predicting factor of non-small cell lung cancer and regulates redox homeostasis
Article Snippet: After the transfer, 5% bovine serum albumin (Thermo Fisher Scientific, Inc.) was used to block the membranes for 1 h at room temperature. .. The following primary antibodies were incubated overnight at 4°C: SPINK1 (1:1,000; catalog no. ab183034; Abcam), β-actin (1:5,000; cat. no. ab8226; Abcam).

Article Title: Single-cell protein profiling in microchambers with barcoded beads
Article Snippet: After incubation, the beads were washed six times with 10 mM PBS. .. Finally, the beads were transferred to a new Eppendorf tube (Eppendorf Protein low-bind tubes, Thermo Fisher Scientific, Waltham, MA, USA) and stored at 4 °C in PBS supplemented with 1% [w/w] bovine serum albumin (BSA, heat shock fraction, Thermo Fisher Scientific, Waltham, MA, USA) until use.

Article Title: ZAR1 and ZAR2 are required for oocyte meiotic maturation by regulating the maternal transcriptome and mRNA translational activation
Article Snippet: .. Immunofluorescence Oocytes were fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS) for 30 min and permeabilized in PBS containing 0.3% Triton X-100 for 20 min. After being blocked with 1% bovine serum albumin in PBS, the oocytes were incubated with primary antibodies for 1 h and sequentially labeled with Alexa Fluor 594- or 488-conjugated secondary antibodies (Molecular Probes) and 4′,6-diamidino-2-phenylindole (DAPI, Vector Laboratories) for 30 min. Oocytes were imaged using a Zeiss LSM710 confocal microscope. .. Detection of transcription and protein synthesis in oocytes To detect transcriptional activity, oocytes were cultured in M16 medium containing 1 mM 5-ethynyl uridine (EU) for 1 h. EU staining was performed using a Click-iT® RNA Alexa Fluor® 488 Imaging Kit (Life Technologies) according to the manufacturer's instructions.

Article Title: Systematic Phosphorylation Analysis of Human Mitotic Protein Complexes
Article Snippet: .. Coverslips were incubated for 1 hr each at room temperature with primary and secondary antibodies (at 2 μg/ml in 3% bovine serum albumin (BSA) in PBS containing 0.01% Triton X-100), DNA was counterstained with Hoechst 33342, and samples were mounted with either ProLong Gold (Molecular Probes / Invitrogen) or Vectashield Mounting Medium (H1000, Vector Laboratories) onto slides. .. Image acquisition was performed as described ( ) or (for BI 4834 characterization) images were taken on a Zeiss Axioplan 2 microscope using a 100× Plan-Apochromat objective lens (Carl Zeiss, Jena, Germany) and equipped with a CoolSnap HQ chargecoupled device camera (Photometrics).

Bradford Assay:

Article Title: Delayed Dark Adaptation in 11-cis-Retinol Dehydrogenase-deficient Mice
Article Snippet: .. Protein concentration was determined by the Bradford assay (Bio-Rad) using bovine serum albumin (Pierce) as the standard. ..

Western Blot:

Article Title: SPINK1 is a prognosis predicting factor of non-small cell lung cancer and regulates redox homeostasis
Article Snippet: Paragraph title: Western blot analysis ... After the transfer, 5% bovine serum albumin (Thermo Fisher Scientific, Inc.) was used to block the membranes for 1 h at room temperature.

Flow Cytometry:

Article Title: Astragalus polysaccharides (PG2) Enhances the M1 Polarization of Macrophages, Functional Maturation of Dendritic Cells, and T Cell-Mediated Anticancer Immune Responses in Patients with Lung Cancer
Article Snippet: .. Flow Cytometry Analyses After staining the THP-1 cells in an Invitrogen LIVE/DEAD Fixable Red Dead Cell Stain kit (#L23102, Thermo Fisher Scientific Inc., Waltham, MA, USA), viable cells were blocked using 10μg/mL human immunoglobulin G (IgG) diluted in 0.5% bovine serum albumin (BSA)/1mM, ethylenediaminetetraacetic acid (EDTA)/0.01%, sodium azide staining buffer on ice for 10 min, and then incubated with anti-human CD206 (MMR)-PE antibody (#12-2069-41, 1:50, eBioscience, Thermo Fisher Scientific Inc., Waltham, MA, USA) on ice for 1 h, before cell fixation in 2% paraformaldehyde (PFA) on ice for 15 min. Next, the cells were incubated with anti-human CD80 (B7-1) Fluorescein isothiocyanate (FITC) antibodies (#11-0809-41, 1:50, eBioscience, Thermo Fisher Scientific Inc., Waltham, MA, USA) in 0.5% saponin on ice for 45 min, washed with staining buffer, and then flow-cytometry data acquisition was performed using the BD FACSCanto II system (BD Biosciences, Thermo Fisher Scientific Inc. Waltham, MA, USA). .. Data obtained were analyzed using Flowjo software v. 9.6.2 (FlowJo, LLC, Tree Star, Inc., Ashland, OR, USA).

Article Title: Enzyme-responsive nanocomposites for wound infection prophylaxis in burn management: in vitro evaluation of their compatibility with healing processes
Article Snippet: Paragraph title: Uptake: microscopic and flow cytometric cell analysis ... Subsequently, secondary antibody (Alexa Fluor 488 and 594, Life Technologies, Carlsbad, CA, USA, 1:1,000 in 1% bovine serum albumin in PBS) was added and incubated for 60 minutes.

Transfection:

Article Title: Vesicular Stomatitis Virus Transcription Is Inhibited by TRIM69 in the Interferon-Induced Antiviral State
Article Snippet: Sixteen hours later, cells were transfected or not with pCAGGS-eGFP/P and pCAGGS-eGFP/P(E67G) plasmids with Lipofectamine 2000 (catalog number 11668-019; Invitrogen), following the manufacturer’s protocol. .. Twenty-four hours posttransfection, cells were washed with 500 μl Dulbecco’s PBS (DPBS) (catalog number 59300C; Sigma), fixed for 15 min at room temperature with 250 μl DPBS containing 2% paraformaldehyde, washed twice with 1 ml DPBS containing 10 mM glycine (to quench residual paraformaldehyde), stained for 15 min with DAPI (4′,6-diamidino-2-phenylindole) diluted in DPBS containing 0.5% bovine serum albumin, and washed twice with 1 ml DPBS containing 0.05% Tween 20 and once with 1 ml H2 O. Coverslips were mounted onto slides with 4 μl ProlongGold (catalog number P36930; Invitrogen).

Dissection:

Article Title: A sperm peptide enhances long-term memory in female Drosophila
Article Snippet: Before dissection, whole F1 female flies (3 to 4 days after eclosion at 25°C) were fixed in 4% paraformaldehyde in PBT (phosphate-buffered saline containing 1% Triton X-100) overnight at 4°C. .. Samples were then rinsed three times for 20 min in PBT, blocked with 2% bovine serum albumin in PBT for 2 hours, and incubated with rabbit anti-GFP (1:400; Invitrogen Molecular Probes) and mouse anti-nc82 (1:100; Developmental Studies Hybridoma Bank) primary antibodies in the blocking solution overnight at 4°C.

Generated:

Article Title: Delayed Dark Adaptation in 11-cis-Retinol Dehydrogenase-deficient Mice
Article Snippet: Anti-RDH11 polyclonal antibody was generated against the C-terminal peptide LWDVSCDLLGLPVDW conjugated to keyhole limpet hemocyanin by Genemed Synthesis Inc. (San Francisco, CA). .. Protein concentration was determined by the Bradford assay (Bio-Rad) using bovine serum albumin (Pierce) as the standard.

Inverted Microscopy:

Article Title: Vesicular Stomatitis Virus Transcription Is Inhibited by TRIM69 in the Interferon-Induced Antiviral State
Article Snippet: Twenty-four hours posttransfection, cells were washed with 500 μl Dulbecco’s PBS (DPBS) (catalog number 59300C; Sigma), fixed for 15 min at room temperature with 250 μl DPBS containing 2% paraformaldehyde, washed twice with 1 ml DPBS containing 10 mM glycine (to quench residual paraformaldehyde), stained for 15 min with DAPI (4′,6-diamidino-2-phenylindole) diluted in DPBS containing 0.5% bovine serum albumin, and washed twice with 1 ml DPBS containing 0.05% Tween 20 and once with 1 ml H2 O. Coverslips were mounted onto slides with 4 μl ProlongGold (catalog number P36930; Invitrogen). .. Confocal images were acquired on a Nikon T1 inverted microscope equipped with a Yokogawa CSU-W1 scan head, a Toptica laser launch, and an Andor Zyla 4.2 plus sCMOS (scientific complementary metal-oxide semiconductor) camera using a plan apochromat lambda 100×/1.45 numeric aperture (NA) differential inference contrast (DIC) oil objective.

Protein Concentration:

Article Title: Delayed Dark Adaptation in 11-cis-Retinol Dehydrogenase-deficient Mice
Article Snippet: .. Protein concentration was determined by the Bradford assay (Bio-Rad) using bovine serum albumin (Pierce) as the standard. ..

Article Title: SPINK1 is a prognosis predicting factor of non-small cell lung cancer and regulates redox homeostasis
Article Snippet: Protein concentration was determined using a bicinchoninic acid assay kit (Beyotime institute of Biotechnology). .. After the transfer, 5% bovine serum albumin (Thermo Fisher Scientific, Inc.) was used to block the membranes for 1 h at room temperature.

Sonication:

Article Title: Topology and Boundaries of the Aerotaxis Receptor Aer in the Membrane of Escherichia coli
Article Snippet: Membrane pellets were resuspended by sonication in high-salt buffer 1 (20 mM sodium phosphate [pH 7.0], 2 M KCl, 10% glycerol, 10 mM EDTA, 1 mM 1,10 phenanthroline containing freshly added 5 mM DTT, and 2 mM PEFAbloc), pelleted, resuspended in high-salt buffer 2 (without DTT or 1,10-phenanthroline), pelleted, resuspended in final buffer (with no DTT; 1,10-phenanthroline; or KCl), pelleted, resuspended in 200 μl of final buffer, aliquoted, frozen in a dry ice-ethanol bath, and stored at −80°C. .. The concentration of protein in the membrane was measured by a BCA (bicinchoninic acid) assay (Pierce Chemical) using bovine serum albumin as the standard.

Binding Assay:

Article Title: How does conserved dopamine neurotrophic factor protect against and rescue neurodegeneration of PC12 cells?
Article Snippet: Cells were washed three times with gentle shaking in 0.1% bovine serum albumin (Beyotime Institute of Biotechnology, Shanghai, China) in PBS, and then incubated with primary antibodies diluted in 0.1% bovine serum albumin/PBS (TH, 1:500; and alpha-synuclein, 1:200; anti mouse monoclonal antibodies; Invitrogen, Paisley, UK) at 4°C overnight. .. Specific antibody binding was detected by Alexa Fluor 488 (green label)-conjugated ExtrAvidin (Sigma-Aldrich Corp.).

Immunofluorescence:

Article Title: How does conserved dopamine neurotrophic factor protect against and rescue neurodegeneration of PC12 cells?
Article Snippet: Paragraph title: Immunofluorescence staining for tyrosine hydroxylase (TH) and alpha-synuclein ... Cells were washed three times with gentle shaking in 0.1% bovine serum albumin (Beyotime Institute of Biotechnology, Shanghai, China) in PBS, and then incubated with primary antibodies diluted in 0.1% bovine serum albumin/PBS (TH, 1:500; and alpha-synuclein, 1:200; anti mouse monoclonal antibodies; Invitrogen, Paisley, UK) at 4°C overnight.

Article Title: ZAR1 and ZAR2 are required for oocyte meiotic maturation by regulating the maternal transcriptome and mRNA translational activation
Article Snippet: .. Immunofluorescence Oocytes were fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS) for 30 min and permeabilized in PBS containing 0.3% Triton X-100 for 20 min. After being blocked with 1% bovine serum albumin in PBS, the oocytes were incubated with primary antibodies for 1 h and sequentially labeled with Alexa Fluor 594- or 488-conjugated secondary antibodies (Molecular Probes) and 4′,6-diamidino-2-phenylindole (DAPI, Vector Laboratories) for 30 min. Oocytes were imaged using a Zeiss LSM710 confocal microscope. .. Detection of transcription and protein synthesis in oocytes To detect transcriptional activity, oocytes were cultured in M16 medium containing 1 mM 5-ethynyl uridine (EU) for 1 h. EU staining was performed using a Click-iT® RNA Alexa Fluor® 488 Imaging Kit (Life Technologies) according to the manufacturer's instructions.

Article Title: Systematic Phosphorylation Analysis of Human Mitotic Protein Complexes
Article Snippet: Paragraph title: Immunofluorescence microscopy ... Coverslips were incubated for 1 hr each at room temperature with primary and secondary antibodies (at 2 μg/ml in 3% bovine serum albumin (BSA) in PBS containing 0.01% Triton X-100), DNA was counterstained with Hoechst 33342, and samples were mounted with either ProLong Gold (Molecular Probes / Invitrogen) or Vectashield Mounting Medium (H1000, Vector Laboratories) onto slides.

Fluorescence:

Article Title: How does conserved dopamine neurotrophic factor protect against and rescue neurodegeneration of PC12 cells?
Article Snippet: Cells were washed three times with gentle shaking in 0.1% bovine serum albumin (Beyotime Institute of Biotechnology, Shanghai, China) in PBS, and then incubated with primary antibodies diluted in 0.1% bovine serum albumin/PBS (TH, 1:500; and alpha-synuclein, 1:200; anti mouse monoclonal antibodies; Invitrogen, Paisley, UK) at 4°C overnight. .. Fluorescence density was quantified.

Article Title: Enzyme-responsive nanocomposites for wound infection prophylaxis in burn management: in vitro evaluation of their compatibility with healing processes
Article Snippet: Subsequently, secondary antibody (Alexa Fluor 488 and 594, Life Technologies, Carlsbad, CA, USA, 1:1,000 in 1% bovine serum albumin in PBS) was added and incubated for 60 minutes. .. Processing of images and quantification of fluorescence and nuclei were performed using the open source software ImageJ 1.47n (National Institutes of Health, Bethesda, MD, USA).

Antibody Labeling:

Article Title: Delayed Dark Adaptation in 11-cis-Retinol Dehydrogenase-deficient Mice
Article Snippet: After washing membranes with PBST, blots were incubated with goat anti-rabbit IgG conjugated to horseradish peroxidase (Pierce) at 1:10,000 dilution, and antibody labeling was detected by chemiluminescence (Pierce). .. Protein concentration was determined by the Bradford assay (Bio-Rad) using bovine serum albumin (Pierce) as the standard.

Immunohistochemistry:

Article Title: A sperm peptide enhances long-term memory in female Drosophila
Article Snippet: Paragraph title: Immunohistochemistry ... Samples were then rinsed three times for 20 min in PBT, blocked with 2% bovine serum albumin in PBT for 2 hours, and incubated with rabbit anti-GFP (1:400; Invitrogen Molecular Probes) and mouse anti-nc82 (1:100; Developmental Studies Hybridoma Bank) primary antibodies in the blocking solution overnight at 4°C.

Microscopy:

Article Title: A sperm peptide enhances long-term memory in female Drosophila
Article Snippet: Samples were then rinsed three times for 20 min in PBT, blocked with 2% bovine serum albumin in PBT for 2 hours, and incubated with rabbit anti-GFP (1:400; Invitrogen Molecular Probes) and mouse anti-nc82 (1:100; Developmental Studies Hybridoma Bank) primary antibodies in the blocking solution overnight at 4°C. .. After three washes (20 min), brains were mounted in ProLong Mounting Medium (Life Technologies) for microscopy analysis.

Article Title: ZAR1 and ZAR2 are required for oocyte meiotic maturation by regulating the maternal transcriptome and mRNA translational activation
Article Snippet: .. Immunofluorescence Oocytes were fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS) for 30 min and permeabilized in PBS containing 0.3% Triton X-100 for 20 min. After being blocked with 1% bovine serum albumin in PBS, the oocytes were incubated with primary antibodies for 1 h and sequentially labeled with Alexa Fluor 594- or 488-conjugated secondary antibodies (Molecular Probes) and 4′,6-diamidino-2-phenylindole (DAPI, Vector Laboratories) for 30 min. Oocytes were imaged using a Zeiss LSM710 confocal microscope. .. Detection of transcription and protein synthesis in oocytes To detect transcriptional activity, oocytes were cultured in M16 medium containing 1 mM 5-ethynyl uridine (EU) for 1 h. EU staining was performed using a Click-iT® RNA Alexa Fluor® 488 Imaging Kit (Life Technologies) according to the manufacturer's instructions.

Article Title: Systematic Phosphorylation Analysis of Human Mitotic Protein Complexes
Article Snippet: Paragraph title: Immunofluorescence microscopy ... Coverslips were incubated for 1 hr each at room temperature with primary and secondary antibodies (at 2 μg/ml in 3% bovine serum albumin (BSA) in PBS containing 0.01% Triton X-100), DNA was counterstained with Hoechst 33342, and samples were mounted with either ProLong Gold (Molecular Probes / Invitrogen) or Vectashield Mounting Medium (H1000, Vector Laboratories) onto slides.

Protein Extraction:

Article Title: Delayed Dark Adaptation in 11-cis-Retinol Dehydrogenase-deficient Mice
Article Snippet: Mouse testes were solubilized in 6 volumes (w/v) of T-PER tissue protein extraction buffer (Pierce Biotechnology). .. Protein concentration was determined by the Bradford assay (Bio-Rad) using bovine serum albumin (Pierce) as the standard.

Labeling:

Article Title: Delayed Dark Adaptation in 11-cis-Retinol Dehydrogenase-deficient Mice
Article Snippet: For immunoblotting, membranes were blocked in PBS containing 0.1% Tween (PBST) and 5% milk (PBSTM) and labeled with an anti-RDH11 polyclonal antibody (2479) diluted to 1:5000 in PBSTM. .. Protein concentration was determined by the Bradford assay (Bio-Rad) using bovine serum albumin (Pierce) as the standard.

Article Title: How does conserved dopamine neurotrophic factor protect against and rescue neurodegeneration of PC12 cells?
Article Snippet: Cells were washed three times with gentle shaking in 0.1% bovine serum albumin (Beyotime Institute of Biotechnology, Shanghai, China) in PBS, and then incubated with primary antibodies diluted in 0.1% bovine serum albumin/PBS (TH, 1:500; and alpha-synuclein, 1:200; anti mouse monoclonal antibodies; Invitrogen, Paisley, UK) at 4°C overnight. .. Labeled donkey anti-rabbit immunoglobulin G (IgG) (1:1,000; Invitrogen) was used as the secondary antibody, with the solution incubated in the dark for 2 hours at room temperature.

Article Title: ZAR1 and ZAR2 are required for oocyte meiotic maturation by regulating the maternal transcriptome and mRNA translational activation
Article Snippet: .. Immunofluorescence Oocytes were fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS) for 30 min and permeabilized in PBS containing 0.3% Triton X-100 for 20 min. After being blocked with 1% bovine serum albumin in PBS, the oocytes were incubated with primary antibodies for 1 h and sequentially labeled with Alexa Fluor 594- or 488-conjugated secondary antibodies (Molecular Probes) and 4′,6-diamidino-2-phenylindole (DAPI, Vector Laboratories) for 30 min. Oocytes were imaged using a Zeiss LSM710 confocal microscope. .. Detection of transcription and protein synthesis in oocytes To detect transcriptional activity, oocytes were cultured in M16 medium containing 1 mM 5-ethynyl uridine (EU) for 1 h. EU staining was performed using a Click-iT® RNA Alexa Fluor® 488 Imaging Kit (Life Technologies) according to the manufacturer's instructions.

Confocal Microscopy:

Article Title: Vesicular Stomatitis Virus Transcription Is Inhibited by TRIM69 in the Interferon-Induced Antiviral State
Article Snippet: Paragraph title: Confocal microscopy. ... Twenty-four hours posttransfection, cells were washed with 500 μl Dulbecco’s PBS (DPBS) (catalog number 59300C; Sigma), fixed for 15 min at room temperature with 250 μl DPBS containing 2% paraformaldehyde, washed twice with 1 ml DPBS containing 10 mM glycine (to quench residual paraformaldehyde), stained for 15 min with DAPI (4′,6-diamidino-2-phenylindole) diluted in DPBS containing 0.5% bovine serum albumin, and washed twice with 1 ml DPBS containing 0.05% Tween 20 and once with 1 ml H2 O. Coverslips were mounted onto slides with 4 μl ProlongGold (catalog number P36930; Invitrogen).

SDS Page:

Article Title: SPINK1 is a prognosis predicting factor of non-small cell lung cancer and regulates redox homeostasis
Article Snippet: Protein (20 µg) were separated by 10% SDS-PAGE and transferred onto a nitrocellulose membrane. .. After the transfer, 5% bovine serum albumin (Thermo Fisher Scientific, Inc.) was used to block the membranes for 1 h at room temperature.

Software:

Article Title: Astragalus polysaccharides (PG2) Enhances the M1 Polarization of Macrophages, Functional Maturation of Dendritic Cells, and T Cell-Mediated Anticancer Immune Responses in Patients with Lung Cancer
Article Snippet: Flow Cytometry Analyses After staining the THP-1 cells in an Invitrogen LIVE/DEAD Fixable Red Dead Cell Stain kit (#L23102, Thermo Fisher Scientific Inc., Waltham, MA, USA), viable cells were blocked using 10μg/mL human immunoglobulin G (IgG) diluted in 0.5% bovine serum albumin (BSA)/1mM, ethylenediaminetetraacetic acid (EDTA)/0.01%, sodium azide staining buffer on ice for 10 min, and then incubated with anti-human CD206 (MMR)-PE antibody (#12-2069-41, 1:50, eBioscience, Thermo Fisher Scientific Inc., Waltham, MA, USA) on ice for 1 h, before cell fixation in 2% paraformaldehyde (PFA) on ice for 15 min. Next, the cells were incubated with anti-human CD80 (B7-1) Fluorescein isothiocyanate (FITC) antibodies (#11-0809-41, 1:50, eBioscience, Thermo Fisher Scientific Inc., Waltham, MA, USA) in 0.5% saponin on ice for 45 min, washed with staining buffer, and then flow-cytometry data acquisition was performed using the BD FACSCanto II system (BD Biosciences, Thermo Fisher Scientific Inc. Waltham, MA, USA). .. Data obtained were analyzed using Flowjo software v. 9.6.2 (FlowJo, LLC, Tree Star, Inc., Ashland, OR, USA).

Article Title: Enzyme-responsive nanocomposites for wound infection prophylaxis in burn management: in vitro evaluation of their compatibility with healing processes
Article Snippet: Subsequently, secondary antibody (Alexa Fluor 488 and 594, Life Technologies, Carlsbad, CA, USA, 1:1,000 in 1% bovine serum albumin in PBS) was added and incubated for 60 minutes. .. Processing of images and quantification of fluorescence and nuclei were performed using the open source software ImageJ 1.47n (National Institutes of Health, Bethesda, MD, USA).

Article Title: Vesicular Stomatitis Virus Transcription Is Inhibited by TRIM69 in the Interferon-Induced Antiviral State
Article Snippet: Twenty-four hours posttransfection, cells were washed with 500 μl Dulbecco’s PBS (DPBS) (catalog number 59300C; Sigma), fixed for 15 min at room temperature with 250 μl DPBS containing 2% paraformaldehyde, washed twice with 1 ml DPBS containing 10 mM glycine (to quench residual paraformaldehyde), stained for 15 min with DAPI (4′,6-diamidino-2-phenylindole) diluted in DPBS containing 0.5% bovine serum albumin, and washed twice with 1 ml DPBS containing 0.05% Tween 20 and once with 1 ml H2 O. Coverslips were mounted onto slides with 4 μl ProlongGold (catalog number P36930; Invitrogen). .. The acquisition software was NIS Elements AR 5.02.

Article Title: Systematic Phosphorylation Analysis of Human Mitotic Protein Complexes
Article Snippet: Coverslips were incubated for 1 hr each at room temperature with primary and secondary antibodies (at 2 μg/ml in 3% bovine serum albumin (BSA) in PBS containing 0.01% Triton X-100), DNA was counterstained with Hoechst 33342, and samples were mounted with either ProLong Gold (Molecular Probes / Invitrogen) or Vectashield Mounting Medium (H1000, Vector Laboratories) onto slides. .. Images were processed using ImageJ software ( ) ( ).

Acid Assay:

Article Title: SPINK1 is a prognosis predicting factor of non-small cell lung cancer and regulates redox homeostasis
Article Snippet: Protein concentration was determined using a bicinchoninic acid assay kit (Beyotime institute of Biotechnology). .. After the transfer, 5% bovine serum albumin (Thermo Fisher Scientific, Inc.) was used to block the membranes for 1 h at room temperature.

Article Title: Topology and Boundaries of the Aerotaxis Receptor Aer in the Membrane of Escherichia coli
Article Snippet: .. The concentration of protein in the membrane was measured by a BCA (bicinchoninic acid) assay (Pierce Chemical) using bovine serum albumin as the standard. .. Frozen membrane preparations were diluted to the required concentration with final buffer prior to use.

Concentration Assay:

Article Title: Topology and Boundaries of the Aerotaxis Receptor Aer in the Membrane of Escherichia coli
Article Snippet: .. The concentration of protein in the membrane was measured by a BCA (bicinchoninic acid) assay (Pierce Chemical) using bovine serum albumin as the standard. .. Frozen membrane preparations were diluted to the required concentration with final buffer prior to use.

Article Title: Single-cell protein profiling in microchambers with barcoded beads
Article Snippet: To allow for selective cell labelling, magnetic particles of different sizes were washed three times on a custom magnetic rack in 10 mM phosphate-buffered saline (PBS, Sigma-Aldrich, St. Louis, MO, USA) and diluted to a final concentration of 0.2 mg mL−1 . .. Finally, the beads were transferred to a new Eppendorf tube (Eppendorf Protein low-bind tubes, Thermo Fisher Scientific, Waltham, MA, USA) and stored at 4 °C in PBS supplemented with 1% [w/w] bovine serum albumin (BSA, heat shock fraction, Thermo Fisher Scientific, Waltham, MA, USA) until use.

Staining:

Article Title: Vascular endothelial growth factor improves the therapeutic effects of cyclodextrin in Niemann-Pick type C mice
Article Snippet: .. Immunofluorescence staining For immunofluorescence staining, brain sections were blocked with phosphate buffered saline (PBS) containing 5% normal goat serum (Vector Laboratories), 2% bovine serum albumin (BSA; Gibco) and 0.4% Triton X-100 (Sigma-Aldrich). .. Sections were then incubated for 24 h with primary antibodies in the same buffer solution.

Article Title: How does conserved dopamine neurotrophic factor protect against and rescue neurodegeneration of PC12 cells?
Article Snippet: Paragraph title: Immunofluorescence staining for tyrosine hydroxylase (TH) and alpha-synuclein ... Cells were washed three times with gentle shaking in 0.1% bovine serum albumin (Beyotime Institute of Biotechnology, Shanghai, China) in PBS, and then incubated with primary antibodies diluted in 0.1% bovine serum albumin/PBS (TH, 1:500; and alpha-synuclein, 1:200; anti mouse monoclonal antibodies; Invitrogen, Paisley, UK) at 4°C overnight.

Article Title: Astragalus polysaccharides (PG2) Enhances the M1 Polarization of Macrophages, Functional Maturation of Dendritic Cells, and T Cell-Mediated Anticancer Immune Responses in Patients with Lung Cancer
Article Snippet: .. Flow Cytometry Analyses After staining the THP-1 cells in an Invitrogen LIVE/DEAD Fixable Red Dead Cell Stain kit (#L23102, Thermo Fisher Scientific Inc., Waltham, MA, USA), viable cells were blocked using 10μg/mL human immunoglobulin G (IgG) diluted in 0.5% bovine serum albumin (BSA)/1mM, ethylenediaminetetraacetic acid (EDTA)/0.01%, sodium azide staining buffer on ice for 10 min, and then incubated with anti-human CD206 (MMR)-PE antibody (#12-2069-41, 1:50, eBioscience, Thermo Fisher Scientific Inc., Waltham, MA, USA) on ice for 1 h, before cell fixation in 2% paraformaldehyde (PFA) on ice for 15 min. Next, the cells were incubated with anti-human CD80 (B7-1) Fluorescein isothiocyanate (FITC) antibodies (#11-0809-41, 1:50, eBioscience, Thermo Fisher Scientific Inc., Waltham, MA, USA) in 0.5% saponin on ice for 45 min, washed with staining buffer, and then flow-cytometry data acquisition was performed using the BD FACSCanto II system (BD Biosciences, Thermo Fisher Scientific Inc. Waltham, MA, USA). .. Data obtained were analyzed using Flowjo software v. 9.6.2 (FlowJo, LLC, Tree Star, Inc., Ashland, OR, USA).

Article Title: Vesicular Stomatitis Virus Transcription Is Inhibited by TRIM69 in the Interferon-Induced Antiviral State
Article Snippet: .. Twenty-four hours posttransfection, cells were washed with 500 μl Dulbecco’s PBS (DPBS) (catalog number 59300C; Sigma), fixed for 15 min at room temperature with 250 μl DPBS containing 2% paraformaldehyde, washed twice with 1 ml DPBS containing 10 mM glycine (to quench residual paraformaldehyde), stained for 15 min with DAPI (4′,6-diamidino-2-phenylindole) diluted in DPBS containing 0.5% bovine serum albumin, and washed twice with 1 ml DPBS containing 0.05% Tween 20 and once with 1 ml H2 O. Coverslips were mounted onto slides with 4 μl ProlongGold (catalog number P36930; Invitrogen). .. Confocal images were acquired on a Nikon T1 inverted microscope equipped with a Yokogawa CSU-W1 scan head, a Toptica laser launch, and an Andor Zyla 4.2 plus sCMOS (scientific complementary metal-oxide semiconductor) camera using a plan apochromat lambda 100×/1.45 numeric aperture (NA) differential inference contrast (DIC) oil objective.

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    Thermo Fisher bsa v nps
    Cellular uptake of free DOX and <t>DOX-BSA-V-NPs.</t> Notes: Cellular internalization of free DOX and DOX-BSA-V-NPs observed by ( A , B ) inverted fluorescence microscopy (100× magnification), ( C – F ) flow cytometry, and ( G ) CLSM (400× magnification). ( C ) 1, control; 2, free DOX (0.78 μg/mL, 2 hours); 3, DOX-BSA-V-NPs (0.78 μg/mL, 2 hours); 4, free DOX (6.24 μg/mL, 2 hours); 5, DOX-BSA-V-NPs (6.24 μg/mL, 2 hours). ( D ) 1′, control; 2′ free DOX (1.56 μg/mL, 0.5 hours); 3′, DOX-BSA-V-NPs (1.56 μg/mL, 0.5 hours); 4′, free DOX (1.56 μg/mL, 8 hours); 5′, DOX-BSA-V-NPs (1.56 μg/mL, 8 hours). ( E ) Concentration-dependent uptake of free DOX and DOX-BSA-V-NPs. The cells were exposed to various concentrations of the DOX formulations at 37°C for 4 hours, and subsequently determined by flow cytometry. ( F ) Time-dependent uptake of free DOX and DOX-BSA-V-NPs. The cells were treated with the DOX formulations at a concentration of 1.56 μg/mL at 37°C and then analyzed by flow cytometry. ( G ) DOX (1.56 μg/mL, 4 hours) and DOX-BSA-V-NPs (1.56 μg/mL, 4 hours). Abbreviations: BSA, bovine serum albumin; CLSM, confocal laser scanning microscopy; DAPI, 4′,6-diamidino-2-phenylindole; DOX, doxorubicin; NPs, nanoparticles; V, vanillin.
    Bsa V Nps, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    bsa v nps - by Bioz Stars, 2020-04
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    86
    Thermo Fisher bsa cuse nanosnakes
    TEM images of <t>BSA–CuSe</t> <t>nanosnakes</t> obtained after different aging time in the typical experiment: a 24 h, b 48 h, and c 96 h, respectively. d HRTEM image of an individual nanosnake. e SAED pattern in an area including many nanosnakes. f The histogram of nanosnakes at 96 h
    Bsa Cuse Nanosnakes, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bsa cuse nanosnakes/product/Thermo Fisher
    Average 86 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    bsa cuse nanosnakes - by Bioz Stars, 2020-04
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    86
    Thermo Fisher control unglycated bsa
    Effect of glyceraldehyde-derived advanced glycation end-products-treated CM on human umbilical vein endothelial cells proliferation. Cell viability was determined with the WST-8 assay. Human umbilical vein endothelial cells were incubated with control <t>unglycated</t> bovine serum albumin <t>(BSA),</t> glyceraldehyde-derived advanced glycation end-products (Glycer-AGEs) (100 μg/mL), CM-BSA, or CM-Glycer-AGEs for 72 h. The open and filled bars represent results for cells treated with control unglycated BSA and Glycer-AGEs, respectively, and the light grey and the black grey bars represent results for cells treated with CM-BSA and CM-Glycer-AGEs, respectively. Data are shown as the mean ± SD ( n = 6), b P
    Control Unglycated Bsa, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/control unglycated bsa/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    control unglycated bsa - by Bioz Stars, 2020-04
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    95
    Thermo Fisher biotinylated bsa
    Principle of the dual nanoparticle-based lateral flow biosensor for simultaneous detection of nodavirus SJNNV and RGNNV genotypes. Two test zones (anti-fluorescein antibody (TZ-R) and anti-digoxigenin antibody (TZ-S)) and a control zone <t>(biotinylated</t> <t>BSA</t> (CZ)) have been deposited on the diagnostic membrane. The sample, containing the respective genotype of the target analyte (tetra-primer PCR product with fluorescein (F) for RGNNV genotype or digoxigenin (D) for SJNNV genotype), is hybridized with a biotinylated genotype-specific oligonucleotide probe and applied on the conjugation pad, where functionalized gold nanoparticles (Au) with anti-biotin antibody have already been added. Following that, the biosensor is immersed in the developing buffer, the sample and the nanoparticles are immobilized on the appropriate test zone, and the positive signal is visible by the naked eye, as a red zone. The excess nanoparticles bind to the control zone of the biosensor. The image shows a side view of the lateral flow biosensor. IP: immersion pad; CP: conjugation pad; M: diagnostic membrane; AP: absorbent pad. The assay components are not in scale.
    Biotinylated Bsa, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotinylated bsa/product/Thermo Fisher
    Average 95 stars, based on 2 article reviews
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    biotinylated bsa - by Bioz Stars, 2020-04
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    Image Search Results


    Cellular uptake of free DOX and DOX-BSA-V-NPs. Notes: Cellular internalization of free DOX and DOX-BSA-V-NPs observed by ( A , B ) inverted fluorescence microscopy (100× magnification), ( C – F ) flow cytometry, and ( G ) CLSM (400× magnification). ( C ) 1, control; 2, free DOX (0.78 μg/mL, 2 hours); 3, DOX-BSA-V-NPs (0.78 μg/mL, 2 hours); 4, free DOX (6.24 μg/mL, 2 hours); 5, DOX-BSA-V-NPs (6.24 μg/mL, 2 hours). ( D ) 1′, control; 2′ free DOX (1.56 μg/mL, 0.5 hours); 3′, DOX-BSA-V-NPs (1.56 μg/mL, 0.5 hours); 4′, free DOX (1.56 μg/mL, 8 hours); 5′, DOX-BSA-V-NPs (1.56 μg/mL, 8 hours). ( E ) Concentration-dependent uptake of free DOX and DOX-BSA-V-NPs. The cells were exposed to various concentrations of the DOX formulations at 37°C for 4 hours, and subsequently determined by flow cytometry. ( F ) Time-dependent uptake of free DOX and DOX-BSA-V-NPs. The cells were treated with the DOX formulations at a concentration of 1.56 μg/mL at 37°C and then analyzed by flow cytometry. ( G ) DOX (1.56 μg/mL, 4 hours) and DOX-BSA-V-NPs (1.56 μg/mL, 4 hours). Abbreviations: BSA, bovine serum albumin; CLSM, confocal laser scanning microscopy; DAPI, 4′,6-diamidino-2-phenylindole; DOX, doxorubicin; NPs, nanoparticles; V, vanillin.

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced tumor delivery and antitumor response of doxorubicin-loaded albumin nanoparticles formulated based on a Schiff base

    doi: 10.2147/IJN.S108689

    Figure Lengend Snippet: Cellular uptake of free DOX and DOX-BSA-V-NPs. Notes: Cellular internalization of free DOX and DOX-BSA-V-NPs observed by ( A , B ) inverted fluorescence microscopy (100× magnification), ( C – F ) flow cytometry, and ( G ) CLSM (400× magnification). ( C ) 1, control; 2, free DOX (0.78 μg/mL, 2 hours); 3, DOX-BSA-V-NPs (0.78 μg/mL, 2 hours); 4, free DOX (6.24 μg/mL, 2 hours); 5, DOX-BSA-V-NPs (6.24 μg/mL, 2 hours). ( D ) 1′, control; 2′ free DOX (1.56 μg/mL, 0.5 hours); 3′, DOX-BSA-V-NPs (1.56 μg/mL, 0.5 hours); 4′, free DOX (1.56 μg/mL, 8 hours); 5′, DOX-BSA-V-NPs (1.56 μg/mL, 8 hours). ( E ) Concentration-dependent uptake of free DOX and DOX-BSA-V-NPs. The cells were exposed to various concentrations of the DOX formulations at 37°C for 4 hours, and subsequently determined by flow cytometry. ( F ) Time-dependent uptake of free DOX and DOX-BSA-V-NPs. The cells were treated with the DOX formulations at a concentration of 1.56 μg/mL at 37°C and then analyzed by flow cytometry. ( G ) DOX (1.56 μg/mL, 4 hours) and DOX-BSA-V-NPs (1.56 μg/mL, 4 hours). Abbreviations: BSA, bovine serum albumin; CLSM, confocal laser scanning microscopy; DAPI, 4′,6-diamidino-2-phenylindole; DOX, doxorubicin; NPs, nanoparticles; V, vanillin.

    Article Snippet: The Raman spectra of DOX, BSA-V-NPs, DOX-BSA-V-NPs, and the mixture of DOX and BSA-V-NPs were recorded by a DXR Raman microscope (Thermo Fisher Scientific) equipped with a 780 nm externally stabilized diode laser.

    Techniques: Fluorescence, Microscopy, Flow Cytometry, Cytometry, Confocal Laser Scanning Microscopy, Concentration Assay

    DSC curves. Notes: Differential scanning calorimetric thermograms of ( A ) mixture of DOX and BSA-V-NPs, ( B ) DOX, ( C ) DOX-BSA-V-NPs, and ( D ) BSA-V-NPs. Abbreviations: BSA, bovine serum albumin; DOX, doxorubicin; NPs, nanoparticles; V, vanillin; DSC, differential scanning calorimetric.

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced tumor delivery and antitumor response of doxorubicin-loaded albumin nanoparticles formulated based on a Schiff base

    doi: 10.2147/IJN.S108689

    Figure Lengend Snippet: DSC curves. Notes: Differential scanning calorimetric thermograms of ( A ) mixture of DOX and BSA-V-NPs, ( B ) DOX, ( C ) DOX-BSA-V-NPs, and ( D ) BSA-V-NPs. Abbreviations: BSA, bovine serum albumin; DOX, doxorubicin; NPs, nanoparticles; V, vanillin; DSC, differential scanning calorimetric.

    Article Snippet: The Raman spectra of DOX, BSA-V-NPs, DOX-BSA-V-NPs, and the mixture of DOX and BSA-V-NPs were recorded by a DXR Raman microscope (Thermo Fisher Scientific) equipped with a 780 nm externally stabilized diode laser.

    Techniques:

    Accumulative release of DOX from DOX-BSA-V-NPs in PBS buffer (pH 7.4 and pH 6.5), 10 μM GSH (pH 7.4), and 20 mM GSH (pH 6.5) with the free DOX as a control. * P

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced tumor delivery and antitumor response of doxorubicin-loaded albumin nanoparticles formulated based on a Schiff base

    doi: 10.2147/IJN.S108689

    Figure Lengend Snippet: Accumulative release of DOX from DOX-BSA-V-NPs in PBS buffer (pH 7.4 and pH 6.5), 10 μM GSH (pH 7.4), and 20 mM GSH (pH 6.5) with the free DOX as a control. * P

    Article Snippet: The Raman spectra of DOX, BSA-V-NPs, DOX-BSA-V-NPs, and the mixture of DOX and BSA-V-NPs were recorded by a DXR Raman microscope (Thermo Fisher Scientific) equipped with a 780 nm externally stabilized diode laser.

    Techniques:

    Raman spectra of ( A ) DOX, ( B ) BSA-V-NPs, ( C ) DOX-BSA-V-NPs, and ( D ) mixture of DOX and BSA-V-NPs. Notes: a, protein band; b, C=O stretching vibrations; c, CH2 bending vibrations; d, ring stretching in the substituted phenyl rings; e and g, amido bands, C=O and C–N stretching vibrations; f, C–N stretching vibrations; h, bending vibrations in substituted phenyl rings. Abbreviations: BSA, bovine serum albumin; DOX, doxorubicin; NPs, nanoparticles; V, vanillin.

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced tumor delivery and antitumor response of doxorubicin-loaded albumin nanoparticles formulated based on a Schiff base

    doi: 10.2147/IJN.S108689

    Figure Lengend Snippet: Raman spectra of ( A ) DOX, ( B ) BSA-V-NPs, ( C ) DOX-BSA-V-NPs, and ( D ) mixture of DOX and BSA-V-NPs. Notes: a, protein band; b, C=O stretching vibrations; c, CH2 bending vibrations; d, ring stretching in the substituted phenyl rings; e and g, amido bands, C=O and C–N stretching vibrations; f, C–N stretching vibrations; h, bending vibrations in substituted phenyl rings. Abbreviations: BSA, bovine serum albumin; DOX, doxorubicin; NPs, nanoparticles; V, vanillin.

    Article Snippet: The Raman spectra of DOX, BSA-V-NPs, DOX-BSA-V-NPs, and the mixture of DOX and BSA-V-NPs were recorded by a DXR Raman microscope (Thermo Fisher Scientific) equipped with a 780 nm externally stabilized diode laser.

    Techniques:

    Stability of DOX-BSA-V-NPs in different conditions. Notes: ( A,1 ) 0.9% NaCl solution; ( A,2 ) 5% glucose solution; ( B,1 ) 10 days at 40°C; ( B,2 ) 60 days at 4°C. Abbreviations: BSA, bovine serum albumin; DOX, doxorubicin; NPs, nanoparticles; V, vanillin.

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced tumor delivery and antitumor response of doxorubicin-loaded albumin nanoparticles formulated based on a Schiff base

    doi: 10.2147/IJN.S108689

    Figure Lengend Snippet: Stability of DOX-BSA-V-NPs in different conditions. Notes: ( A,1 ) 0.9% NaCl solution; ( A,2 ) 5% glucose solution; ( B,1 ) 10 days at 40°C; ( B,2 ) 60 days at 4°C. Abbreviations: BSA, bovine serum albumin; DOX, doxorubicin; NPs, nanoparticles; V, vanillin.

    Article Snippet: The Raman spectra of DOX, BSA-V-NPs, DOX-BSA-V-NPs, and the mixture of DOX and BSA-V-NPs were recorded by a DXR Raman microscope (Thermo Fisher Scientific) equipped with a 780 nm externally stabilized diode laser.

    Techniques:

    CD spectra of BSA-V-NPs and DOX-BSA-V-NPs. Notes: ( A ) BSA-V-NPs (0% DOX); ( B ) DOX-BSA-V-NPs (2.5% DOX); and ( C ) DOX-BSA-V-NPs (5% DOX). Abbreviations: BSA, bovine serum albumin; CD, circular dichroism; DOX, doxorubicin; NPs, nanoparticles; V, vanillin.

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced tumor delivery and antitumor response of doxorubicin-loaded albumin nanoparticles formulated based on a Schiff base

    doi: 10.2147/IJN.S108689

    Figure Lengend Snippet: CD spectra of BSA-V-NPs and DOX-BSA-V-NPs. Notes: ( A ) BSA-V-NPs (0% DOX); ( B ) DOX-BSA-V-NPs (2.5% DOX); and ( C ) DOX-BSA-V-NPs (5% DOX). Abbreviations: BSA, bovine serum albumin; CD, circular dichroism; DOX, doxorubicin; NPs, nanoparticles; V, vanillin.

    Article Snippet: The Raman spectra of DOX, BSA-V-NPs, DOX-BSA-V-NPs, and the mixture of DOX and BSA-V-NPs were recorded by a DXR Raman microscope (Thermo Fisher Scientific) equipped with a 780 nm externally stabilized diode laser.

    Techniques:

    HE staining of ( A ) heart and ( B ) tumor (200× magnification). Notes: Heps tumor-bearing mice were randomly divided into four treatment groups and were administered with ( 1 ) normal saline, ( 2 ) free DOX, ( 3 ) DOX-BSA-NPs, and ( 4 ) DOX-BSA-V-NPs. Black arrow in ( A,1 ), myocardial fibers arranged neatly; black arrow in ( A,2 ), moderate focal edema necrosis; black arrow in ( A,3 ) and ( A,4 ), a slight extension of the myocardial fiber gap; red arrow in ( A,2 ), focal band fibrosis; red arrow in ( B,2 ), inflammatory infiltration; black arrow in ( B,2 ), ( B,3 ) and ( B,4 ), cell apoptosis and spotty necrosis. Abbreviations: BSA, bovine serum albumin; DOX, doxorubicin; HE, hematoxylin and eosin; NPs, nanoparticles; V, vanillin.

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced tumor delivery and antitumor response of doxorubicin-loaded albumin nanoparticles formulated based on a Schiff base

    doi: 10.2147/IJN.S108689

    Figure Lengend Snippet: HE staining of ( A ) heart and ( B ) tumor (200× magnification). Notes: Heps tumor-bearing mice were randomly divided into four treatment groups and were administered with ( 1 ) normal saline, ( 2 ) free DOX, ( 3 ) DOX-BSA-NPs, and ( 4 ) DOX-BSA-V-NPs. Black arrow in ( A,1 ), myocardial fibers arranged neatly; black arrow in ( A,2 ), moderate focal edema necrosis; black arrow in ( A,3 ) and ( A,4 ), a slight extension of the myocardial fiber gap; red arrow in ( A,2 ), focal band fibrosis; red arrow in ( B,2 ), inflammatory infiltration; black arrow in ( B,2 ), ( B,3 ) and ( B,4 ), cell apoptosis and spotty necrosis. Abbreviations: BSA, bovine serum albumin; DOX, doxorubicin; HE, hematoxylin and eosin; NPs, nanoparticles; V, vanillin.

    Article Snippet: The Raman spectra of DOX, BSA-V-NPs, DOX-BSA-V-NPs, and the mixture of DOX and BSA-V-NPs were recorded by a DXR Raman microscope (Thermo Fisher Scientific) equipped with a 780 nm externally stabilized diode laser.

    Techniques: Staining, Mouse Assay

    Biodistribution of free DOX, DOX-BSA-NPs and DOX-BSA-V-NPs in ( A ) heart and ( B ) tumor. Notes: Time profiles of DOX accumulated in the ( A ) heart and ( B ) tumor after intravenous injection of different DOX formulations with a DOX dose of 5 mg kg −1 . DOX/heart and DOX/tumor are the ratios of the DOX amount in the heart (μg) and tumor (μg) to the tumor weight (g), respectively. * P

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced tumor delivery and antitumor response of doxorubicin-loaded albumin nanoparticles formulated based on a Schiff base

    doi: 10.2147/IJN.S108689

    Figure Lengend Snippet: Biodistribution of free DOX, DOX-BSA-NPs and DOX-BSA-V-NPs in ( A ) heart and ( B ) tumor. Notes: Time profiles of DOX accumulated in the ( A ) heart and ( B ) tumor after intravenous injection of different DOX formulations with a DOX dose of 5 mg kg −1 . DOX/heart and DOX/tumor are the ratios of the DOX amount in the heart (μg) and tumor (μg) to the tumor weight (g), respectively. * P

    Article Snippet: The Raman spectra of DOX, BSA-V-NPs, DOX-BSA-V-NPs, and the mixture of DOX and BSA-V-NPs were recorded by a DXR Raman microscope (Thermo Fisher Scientific) equipped with a 780 nm externally stabilized diode laser.

    Techniques: Injection

    In vitro cytotoxicity of DOX and DOX-BSA-V-NPs against BGC-823 cells for 48 hours of treatment assessed by using the CCK-8 assay. * P

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced tumor delivery and antitumor response of doxorubicin-loaded albumin nanoparticles formulated based on a Schiff base

    doi: 10.2147/IJN.S108689

    Figure Lengend Snippet: In vitro cytotoxicity of DOX and DOX-BSA-V-NPs against BGC-823 cells for 48 hours of treatment assessed by using the CCK-8 assay. * P

    Article Snippet: The Raman spectra of DOX, BSA-V-NPs, DOX-BSA-V-NPs, and the mixture of DOX and BSA-V-NPs were recorded by a DXR Raman microscope (Thermo Fisher Scientific) equipped with a 780 nm externally stabilized diode laser.

    Techniques: In Vitro, CCK-8 Assay

    NIRF images of the xenograft Heps tumor-bearing mouse after intravenous injection of DiR-BSA-V-NPs. Abbreviations: BSA, bovine serum albumin; DiR, 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindotricarbocyanine iodide; DOX, doxorubicin; NIRF, near-infrared fluorescence; NPs, nanoparticles; V, vanillin.

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced tumor delivery and antitumor response of doxorubicin-loaded albumin nanoparticles formulated based on a Schiff base

    doi: 10.2147/IJN.S108689

    Figure Lengend Snippet: NIRF images of the xenograft Heps tumor-bearing mouse after intravenous injection of DiR-BSA-V-NPs. Abbreviations: BSA, bovine serum albumin; DiR, 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindotricarbocyanine iodide; DOX, doxorubicin; NIRF, near-infrared fluorescence; NPs, nanoparticles; V, vanillin.

    Article Snippet: The Raman spectra of DOX, BSA-V-NPs, DOX-BSA-V-NPs, and the mixture of DOX and BSA-V-NPs were recorded by a DXR Raman microscope (Thermo Fisher Scientific) equipped with a 780 nm externally stabilized diode laser.

    Techniques: Injection, Fluorescence

    Cellular uptake mechanism investigated by ( A ) CLSM and ( B ) inverted fluorescence microscopy. Notes: Uptake pathways of DOX-BSA-V-NPs in BGC-823 cells studied by using ( A ) LysoTracker Green (400× magnification) and ( B ) endocytic inhibitors (100× magnification). ( A,1 ) DAPI; ( A,2 ) RBITC-labeled BSA-V-NPs; ( A,3 ) LysoTracker Green; ( A,4 ) merge. Abbreviations: BSA, bovine serum albumin; CLSM, confocal laser scanning microscopy; DAPI, 4′,6-diamidino-2-phenylindole; DOX, doxorubicin; RBITC, rhodamin B isothiocyanate; NPs, nanoparticles; V, vanillin.

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced tumor delivery and antitumor response of doxorubicin-loaded albumin nanoparticles formulated based on a Schiff base

    doi: 10.2147/IJN.S108689

    Figure Lengend Snippet: Cellular uptake mechanism investigated by ( A ) CLSM and ( B ) inverted fluorescence microscopy. Notes: Uptake pathways of DOX-BSA-V-NPs in BGC-823 cells studied by using ( A ) LysoTracker Green (400× magnification) and ( B ) endocytic inhibitors (100× magnification). ( A,1 ) DAPI; ( A,2 ) RBITC-labeled BSA-V-NPs; ( A,3 ) LysoTracker Green; ( A,4 ) merge. Abbreviations: BSA, bovine serum albumin; CLSM, confocal laser scanning microscopy; DAPI, 4′,6-diamidino-2-phenylindole; DOX, doxorubicin; RBITC, rhodamin B isothiocyanate; NPs, nanoparticles; V, vanillin.

    Article Snippet: The Raman spectra of DOX, BSA-V-NPs, DOX-BSA-V-NPs, and the mixture of DOX and BSA-V-NPs were recorded by a DXR Raman microscope (Thermo Fisher Scientific) equipped with a 780 nm externally stabilized diode laser.

    Techniques: Confocal Laser Scanning Microscopy, Fluorescence, Microscopy, Labeling

    TEM images of the nanoparticles. Notes: ( A ) BSA-V-NPs, scale bar 100 nm; ( B ) DOX-BSA-V-NPs, scale bar 100 nm. Abbreviations: BSA, bovine serum albumin; DOX, doxorubicin; NPs, nanoparticles; TEM, transmission electron microscopy; V, vanillin.

    Journal: International Journal of Nanomedicine

    Article Title: Enhanced tumor delivery and antitumor response of doxorubicin-loaded albumin nanoparticles formulated based on a Schiff base

    doi: 10.2147/IJN.S108689

    Figure Lengend Snippet: TEM images of the nanoparticles. Notes: ( A ) BSA-V-NPs, scale bar 100 nm; ( B ) DOX-BSA-V-NPs, scale bar 100 nm. Abbreviations: BSA, bovine serum albumin; DOX, doxorubicin; NPs, nanoparticles; TEM, transmission electron microscopy; V, vanillin.

    Article Snippet: The Raman spectra of DOX, BSA-V-NPs, DOX-BSA-V-NPs, and the mixture of DOX and BSA-V-NPs were recorded by a DXR Raman microscope (Thermo Fisher Scientific) equipped with a 780 nm externally stabilized diode laser.

    Techniques: Transmission Electron Microscopy, Transmission Assay, Electron Microscopy

    TEM images of BSA–CuSe nanosnakes obtained after different aging time in the typical experiment: a 24 h, b 48 h, and c 96 h, respectively. d HRTEM image of an individual nanosnake. e SAED pattern in an area including many nanosnakes. f The histogram of nanosnakes at 96 h

    Journal: Nanoscale Research Letters

    Article Title: Copper Selenide Nanosnakes: Bovine Serum Albumin-Assisted Room Temperature Controllable Synthesis and Characterization

    doi: 10.1007/s11671-010-9587-0

    Figure Lengend Snippet: TEM images of BSA–CuSe nanosnakes obtained after different aging time in the typical experiment: a 24 h, b 48 h, and c 96 h, respectively. d HRTEM image of an individual nanosnake. e SAED pattern in an area including many nanosnakes. f The histogram of nanosnakes at 96 h

    Article Snippet: Characterization of Synthesized BSA–CuSe Nanosnakes These synthesized BSA–CuSe nanosnakes were characterized by a UNICAM UV300 spectrophotometer (Thermo Spectronic, USA), high-resolution transmission electron microscopy(HR-TEM, Hitachi H-700H, Hitachi, Japan), selected-area electron diffraction, energy dispersive spectroscopy, a PerkinElmer LS55 spectrofluorimeter, Laser Raman spectroscopy, and Fourier transform infrared (FT-IR) spectroscopy (an FTS135 infrared spectrometer from BIO-RAD, United States).

    Techniques: Transmission Electron Microscopy

    Effects of 20 μg/ml BSA–CuSe nanosnakes and CuSe nanocrystals on Human fibroblast cells

    Journal: Nanoscale Research Letters

    Article Title: Copper Selenide Nanosnakes: Bovine Serum Albumin-Assisted Room Temperature Controllable Synthesis and Characterization

    doi: 10.1007/s11671-010-9587-0

    Figure Lengend Snippet: Effects of 20 μg/ml BSA–CuSe nanosnakes and CuSe nanocrystals on Human fibroblast cells

    Article Snippet: Characterization of Synthesized BSA–CuSe Nanosnakes These synthesized BSA–CuSe nanosnakes were characterized by a UNICAM UV300 spectrophotometer (Thermo Spectronic, USA), high-resolution transmission electron microscopy(HR-TEM, Hitachi H-700H, Hitachi, Japan), selected-area electron diffraction, energy dispersive spectroscopy, a PerkinElmer LS55 spectrofluorimeter, Laser Raman spectroscopy, and Fourier transform infrared (FT-IR) spectroscopy (an FTS135 infrared spectrometer from BIO-RAD, United States).

    Techniques:

    a UV–vis absorption spectrum of BSA, BSA–Cu 2+ , BSA–CuSe at 24 h, BSA–CuSe at 48 h, BSA–CuSe at 72 h, BSA–CuSe at 96 h; b The change of absorbance at 190 nm

    Journal: Nanoscale Research Letters

    Article Title: Copper Selenide Nanosnakes: Bovine Serum Albumin-Assisted Room Temperature Controllable Synthesis and Characterization

    doi: 10.1007/s11671-010-9587-0

    Figure Lengend Snippet: a UV–vis absorption spectrum of BSA, BSA–Cu 2+ , BSA–CuSe at 24 h, BSA–CuSe at 48 h, BSA–CuSe at 72 h, BSA–CuSe at 96 h; b The change of absorbance at 190 nm

    Article Snippet: Characterization of Synthesized BSA–CuSe Nanosnakes These synthesized BSA–CuSe nanosnakes were characterized by a UNICAM UV300 spectrophotometer (Thermo Spectronic, USA), high-resolution transmission electron microscopy(HR-TEM, Hitachi H-700H, Hitachi, Japan), selected-area electron diffraction, energy dispersive spectroscopy, a PerkinElmer LS55 spectrofluorimeter, Laser Raman spectroscopy, and Fourier transform infrared (FT-IR) spectroscopy (an FTS135 infrared spectrometer from BIO-RAD, United States).

    Techniques:

    Schematic mechanism of synthesis of CuSe nanosnakes using BSA as soft-template

    Journal: Nanoscale Research Letters

    Article Title: Copper Selenide Nanosnakes: Bovine Serum Albumin-Assisted Room Temperature Controllable Synthesis and Characterization

    doi: 10.1007/s11671-010-9587-0

    Figure Lengend Snippet: Schematic mechanism of synthesis of CuSe nanosnakes using BSA as soft-template

    Article Snippet: Characterization of Synthesized BSA–CuSe Nanosnakes These synthesized BSA–CuSe nanosnakes were characterized by a UNICAM UV300 spectrophotometer (Thermo Spectronic, USA), high-resolution transmission electron microscopy(HR-TEM, Hitachi H-700H, Hitachi, Japan), selected-area electron diffraction, energy dispersive spectroscopy, a PerkinElmer LS55 spectrofluorimeter, Laser Raman spectroscopy, and Fourier transform infrared (FT-IR) spectroscopy (an FTS135 infrared spectrometer from BIO-RAD, United States).

    Techniques:

    TEM images showing oriented attachment of copper selenide nanosnakes in BSA solution for 48 h. a Low-magnification TEM image of sample. b , c TEM images of two different devour stages of copper selenide nanosnakes. HRTEM images of different parts of an individual nanosnake: d the neck, e the body, f the tail, respectively

    Journal: Nanoscale Research Letters

    Article Title: Copper Selenide Nanosnakes: Bovine Serum Albumin-Assisted Room Temperature Controllable Synthesis and Characterization

    doi: 10.1007/s11671-010-9587-0

    Figure Lengend Snippet: TEM images showing oriented attachment of copper selenide nanosnakes in BSA solution for 48 h. a Low-magnification TEM image of sample. b , c TEM images of two different devour stages of copper selenide nanosnakes. HRTEM images of different parts of an individual nanosnake: d the neck, e the body, f the tail, respectively

    Article Snippet: Characterization of Synthesized BSA–CuSe Nanosnakes These synthesized BSA–CuSe nanosnakes were characterized by a UNICAM UV300 spectrophotometer (Thermo Spectronic, USA), high-resolution transmission electron microscopy(HR-TEM, Hitachi H-700H, Hitachi, Japan), selected-area electron diffraction, energy dispersive spectroscopy, a PerkinElmer LS55 spectrofluorimeter, Laser Raman spectroscopy, and Fourier transform infrared (FT-IR) spectroscopy (an FTS135 infrared spectrometer from BIO-RAD, United States).

    Techniques: Transmission Electron Microscopy

    a The FT-IR spectra of ( a ) pure BSA, ( b ) BSA–Cu 2+ , and ( c ) BSA–CuSe in BSA solution for 96 h. b Raman spectra (632.8 nm excitation) of pure BSA, BSA–Cu 2+ , BSA–CuSe in BSA solution for 96 h

    Journal: Nanoscale Research Letters

    Article Title: Copper Selenide Nanosnakes: Bovine Serum Albumin-Assisted Room Temperature Controllable Synthesis and Characterization

    doi: 10.1007/s11671-010-9587-0

    Figure Lengend Snippet: a The FT-IR spectra of ( a ) pure BSA, ( b ) BSA–Cu 2+ , and ( c ) BSA–CuSe in BSA solution for 96 h. b Raman spectra (632.8 nm excitation) of pure BSA, BSA–Cu 2+ , BSA–CuSe in BSA solution for 96 h

    Article Snippet: Characterization of Synthesized BSA–CuSe Nanosnakes These synthesized BSA–CuSe nanosnakes were characterized by a UNICAM UV300 spectrophotometer (Thermo Spectronic, USA), high-resolution transmission electron microscopy(HR-TEM, Hitachi H-700H, Hitachi, Japan), selected-area electron diffraction, energy dispersive spectroscopy, a PerkinElmer LS55 spectrofluorimeter, Laser Raman spectroscopy, and Fourier transform infrared (FT-IR) spectroscopy (an FTS135 infrared spectrometer from BIO-RAD, United States).

    Techniques:

    TEM images of BSA–CuSe nanosnakes ( a ) and the cubic copper selenide nanostructures ( b )

    Journal: Nanoscale Research Letters

    Article Title: Copper Selenide Nanosnakes: Bovine Serum Albumin-Assisted Room Temperature Controllable Synthesis and Characterization

    doi: 10.1007/s11671-010-9587-0

    Figure Lengend Snippet: TEM images of BSA–CuSe nanosnakes ( a ) and the cubic copper selenide nanostructures ( b )

    Article Snippet: Characterization of Synthesized BSA–CuSe Nanosnakes These synthesized BSA–CuSe nanosnakes were characterized by a UNICAM UV300 spectrophotometer (Thermo Spectronic, USA), high-resolution transmission electron microscopy(HR-TEM, Hitachi H-700H, Hitachi, Japan), selected-area electron diffraction, energy dispersive spectroscopy, a PerkinElmer LS55 spectrofluorimeter, Laser Raman spectroscopy, and Fourier transform infrared (FT-IR) spectroscopy (an FTS135 infrared spectrometer from BIO-RAD, United States).

    Techniques: Transmission Electron Microscopy

    The CD spectra of a pure BSA, b BSA–Cu 2+ , and c BSA–CuSe in BSA solution

    Journal: Nanoscale Research Letters

    Article Title: Copper Selenide Nanosnakes: Bovine Serum Albumin-Assisted Room Temperature Controllable Synthesis and Characterization

    doi: 10.1007/s11671-010-9587-0

    Figure Lengend Snippet: The CD spectra of a pure BSA, b BSA–Cu 2+ , and c BSA–CuSe in BSA solution

    Article Snippet: Characterization of Synthesized BSA–CuSe Nanosnakes These synthesized BSA–CuSe nanosnakes were characterized by a UNICAM UV300 spectrophotometer (Thermo Spectronic, USA), high-resolution transmission electron microscopy(HR-TEM, Hitachi H-700H, Hitachi, Japan), selected-area electron diffraction, energy dispersive spectroscopy, a PerkinElmer LS55 spectrofluorimeter, Laser Raman spectroscopy, and Fourier transform infrared (FT-IR) spectroscopy (an FTS135 infrared spectrometer from BIO-RAD, United States).

    Techniques:

    Effect of glyceraldehyde-derived advanced glycation end-products-treated CM on human umbilical vein endothelial cells proliferation. Cell viability was determined with the WST-8 assay. Human umbilical vein endothelial cells were incubated with control unglycated bovine serum albumin (BSA), glyceraldehyde-derived advanced glycation end-products (Glycer-AGEs) (100 μg/mL), CM-BSA, or CM-Glycer-AGEs for 72 h. The open and filled bars represent results for cells treated with control unglycated BSA and Glycer-AGEs, respectively, and the light grey and the black grey bars represent results for cells treated with CM-BSA and CM-Glycer-AGEs, respectively. Data are shown as the mean ± SD ( n = 6), b P

    Journal: World Journal of Gastroenterology : WJG

    Article Title: Glycer-AGEs-RAGE signaling enhances the angiogenic potential of hepatocellular carcinoma by upregulating VEGF expression

    doi: 10.3748/wjg.v18.i15.1781

    Figure Lengend Snippet: Effect of glyceraldehyde-derived advanced glycation end-products-treated CM on human umbilical vein endothelial cells proliferation. Cell viability was determined with the WST-8 assay. Human umbilical vein endothelial cells were incubated with control unglycated bovine serum albumin (BSA), glyceraldehyde-derived advanced glycation end-products (Glycer-AGEs) (100 μg/mL), CM-BSA, or CM-Glycer-AGEs for 72 h. The open and filled bars represent results for cells treated with control unglycated BSA and Glycer-AGEs, respectively, and the light grey and the black grey bars represent results for cells treated with CM-BSA and CM-Glycer-AGEs, respectively. Data are shown as the mean ± SD ( n = 6), b P

    Article Snippet: Cells (1.5×105 cells/mL) were incubated with 10% FBS/DMEM for 24 h. After removing the stopper covering the center of the well, fluorescently-labeled cells were incubated with control unglycated BSA or Glycer-AGEs for 24 h. The number of cells that had migrated to the center was assessed at excitation and emission wavelengths of 485 nm and 530 nm, respectively, using a fluorescence microplate reader (Labsystems Fluoroskan Ascent CF, Type 374; Thermo Fisher Scientific).

    Techniques: Derivative Assay, Incubation

    Effect of glyceraldehyde-derived advanced glycation end-products on the viability of hepatocellular carcinoma cells. Cell viability was determined using the WST-8 assay. Hep3B (A) and HepG2 (B) cells were incubated with control unglycated bovine serum albumin (BSA) or glyceraldehyde-derived advanced glycation end-products (Glycer-AGEs) (100 μg/mL) for 24 h. The open and filled bars represent results for cells treated with control unglycated BSA and Glycer-AGEs, respectively. Data are shown as the mean ± SD ( n = 6).

    Journal: World Journal of Gastroenterology : WJG

    Article Title: Glycer-AGEs-RAGE signaling enhances the angiogenic potential of hepatocellular carcinoma by upregulating VEGF expression

    doi: 10.3748/wjg.v18.i15.1781

    Figure Lengend Snippet: Effect of glyceraldehyde-derived advanced glycation end-products on the viability of hepatocellular carcinoma cells. Cell viability was determined using the WST-8 assay. Hep3B (A) and HepG2 (B) cells were incubated with control unglycated bovine serum albumin (BSA) or glyceraldehyde-derived advanced glycation end-products (Glycer-AGEs) (100 μg/mL) for 24 h. The open and filled bars represent results for cells treated with control unglycated BSA and Glycer-AGEs, respectively. Data are shown as the mean ± SD ( n = 6).

    Article Snippet: Cells (1.5×105 cells/mL) were incubated with 10% FBS/DMEM for 24 h. After removing the stopper covering the center of the well, fluorescently-labeled cells were incubated with control unglycated BSA or Glycer-AGEs for 24 h. The number of cells that had migrated to the center was assessed at excitation and emission wavelengths of 485 nm and 530 nm, respectively, using a fluorescence microplate reader (Labsystems Fluoroskan Ascent CF, Type 374; Thermo Fisher Scientific).

    Techniques: Derivative Assay, Incubation

    Effect of glyceraldehyde-derived advanced glycation end-products on the malignancy of hepatocellular carcinoma cells. Hep3B and HepG2 cells were incubated with control unglycated bovine serum albumin (BSA) or glyceraldehyde-derived advanced glycation end-products (Glycer-AGEs) for 24 h. A: In Hep3B cells, cyclooxygenase-2 (COX-2) mRNA expression levels were analyzed using real-time reverse transcription-polymerase chain reactions, and results were normalized to the β-actin mRNA level ( n = 3), b P

    Journal: World Journal of Gastroenterology : WJG

    Article Title: Glycer-AGEs-RAGE signaling enhances the angiogenic potential of hepatocellular carcinoma by upregulating VEGF expression

    doi: 10.3748/wjg.v18.i15.1781

    Figure Lengend Snippet: Effect of glyceraldehyde-derived advanced glycation end-products on the malignancy of hepatocellular carcinoma cells. Hep3B and HepG2 cells were incubated with control unglycated bovine serum albumin (BSA) or glyceraldehyde-derived advanced glycation end-products (Glycer-AGEs) for 24 h. A: In Hep3B cells, cyclooxygenase-2 (COX-2) mRNA expression levels were analyzed using real-time reverse transcription-polymerase chain reactions, and results were normalized to the β-actin mRNA level ( n = 3), b P

    Article Snippet: Cells (1.5×105 cells/mL) were incubated with 10% FBS/DMEM for 24 h. After removing the stopper covering the center of the well, fluorescently-labeled cells were incubated with control unglycated BSA or Glycer-AGEs for 24 h. The number of cells that had migrated to the center was assessed at excitation and emission wavelengths of 485 nm and 530 nm, respectively, using a fluorescence microplate reader (Labsystems Fluoroskan Ascent CF, Type 374; Thermo Fisher Scientific).

    Techniques: Derivative Assay, Incubation, Expressing

    Effect of glyceraldehyde-derived advanced glycation end-products on the angiogenesis of hepatocellular carcinoma cells. A and B: Hep3B and HepG2 cells were incubated with control unglycated bovine serum albumin (BSA) or glyceraldehyde-derived advanced glycation end-products (Glycer-AGEs) for 24 h. Vascular endothelial growth factor (VEGF) mRNA expression was analyzed using real-time reverse transcription-polymerase chain reactions, and results were normalized to the β-actin mRNA level; C and D: Hep3B and HepG2 cells were incubated with control unglycated BSA or Glycer-AGEs for 24 or 48 h. The conditioned medium was collected, and VEGF expression levels of the cells were determined by enzyme-linked immunosorbent assay. The open and filled bars represent results for cells treated with control unglycated BSA and Glycer-AGEs, respectively. Data are shown as the mean ± SD ( n = 3), b P

    Journal: World Journal of Gastroenterology : WJG

    Article Title: Glycer-AGEs-RAGE signaling enhances the angiogenic potential of hepatocellular carcinoma by upregulating VEGF expression

    doi: 10.3748/wjg.v18.i15.1781

    Figure Lengend Snippet: Effect of glyceraldehyde-derived advanced glycation end-products on the angiogenesis of hepatocellular carcinoma cells. A and B: Hep3B and HepG2 cells were incubated with control unglycated bovine serum albumin (BSA) or glyceraldehyde-derived advanced glycation end-products (Glycer-AGEs) for 24 h. Vascular endothelial growth factor (VEGF) mRNA expression was analyzed using real-time reverse transcription-polymerase chain reactions, and results were normalized to the β-actin mRNA level; C and D: Hep3B and HepG2 cells were incubated with control unglycated BSA or Glycer-AGEs for 24 or 48 h. The conditioned medium was collected, and VEGF expression levels of the cells were determined by enzyme-linked immunosorbent assay. The open and filled bars represent results for cells treated with control unglycated BSA and Glycer-AGEs, respectively. Data are shown as the mean ± SD ( n = 3), b P

    Article Snippet: Cells (1.5×105 cells/mL) were incubated with 10% FBS/DMEM for 24 h. After removing the stopper covering the center of the well, fluorescently-labeled cells were incubated with control unglycated BSA or Glycer-AGEs for 24 h. The number of cells that had migrated to the center was assessed at excitation and emission wavelengths of 485 nm and 530 nm, respectively, using a fluorescence microplate reader (Labsystems Fluoroskan Ascent CF, Type 374; Thermo Fisher Scientific).

    Techniques: Derivative Assay, Incubation, Expressing, Enzyme-linked Immunosorbent Assay

    Principle of the dual nanoparticle-based lateral flow biosensor for simultaneous detection of nodavirus SJNNV and RGNNV genotypes. Two test zones (anti-fluorescein antibody (TZ-R) and anti-digoxigenin antibody (TZ-S)) and a control zone (biotinylated BSA (CZ)) have been deposited on the diagnostic membrane. The sample, containing the respective genotype of the target analyte (tetra-primer PCR product with fluorescein (F) for RGNNV genotype or digoxigenin (D) for SJNNV genotype), is hybridized with a biotinylated genotype-specific oligonucleotide probe and applied on the conjugation pad, where functionalized gold nanoparticles (Au) with anti-biotin antibody have already been added. Following that, the biosensor is immersed in the developing buffer, the sample and the nanoparticles are immobilized on the appropriate test zone, and the positive signal is visible by the naked eye, as a red zone. The excess nanoparticles bind to the control zone of the biosensor. The image shows a side view of the lateral flow biosensor. IP: immersion pad; CP: conjugation pad; M: diagnostic membrane; AP: absorbent pad. The assay components are not in scale.

    Journal: Journal of Analytical Methods in Chemistry

    Article Title: Towards a Dual Lateral Flow Nanobiosensor for Simultaneous Detection of Virus Genotype-Specific PCR Products

    doi: 10.1155/2018/7691014

    Figure Lengend Snippet: Principle of the dual nanoparticle-based lateral flow biosensor for simultaneous detection of nodavirus SJNNV and RGNNV genotypes. Two test zones (anti-fluorescein antibody (TZ-R) and anti-digoxigenin antibody (TZ-S)) and a control zone (biotinylated BSA (CZ)) have been deposited on the diagnostic membrane. The sample, containing the respective genotype of the target analyte (tetra-primer PCR product with fluorescein (F) for RGNNV genotype or digoxigenin (D) for SJNNV genotype), is hybridized with a biotinylated genotype-specific oligonucleotide probe and applied on the conjugation pad, where functionalized gold nanoparticles (Au) with anti-biotin antibody have already been added. Following that, the biosensor is immersed in the developing buffer, the sample and the nanoparticles are immobilized on the appropriate test zone, and the positive signal is visible by the naked eye, as a red zone. The excess nanoparticles bind to the control zone of the biosensor. The image shows a side view of the lateral flow biosensor. IP: immersion pad; CP: conjugation pad; M: diagnostic membrane; AP: absorbent pad. The assay components are not in scale.

    Article Snippet: The zones were formed by loading anti-fluorescein antibody (TZ-R: polyclonal anti-fluorescein antibody; monoclonal anti-fluorescein antibody), anti-digoxigenin antibody (TZ-S: Roche Diagnostics, Mannheim, Germany), and biotinylated BSA (bBSA: Thermo Fisher Scientific Inc., Rockford, IL, USA) on the membrane and were located at 10, 15, and 20 mm distance from the edge of the membrane, respectively.

    Techniques: Flow Cytometry, Diagnostic Assay, Polymerase Chain Reaction, Conjugation Assay