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  • 99
    Name:
    Bovine Serum Albumin
    Description:

    Catalog Number:
    a9306
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    Structured Review

    Millipore bsa
    Bovine Serum Albumin

    https://www.bioz.com/result/bsa/product/Millipore
    Average 99 stars, based on 1377 article reviews
    Price from $9.99 to $1999.99
    bsa - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Albumin-induced podocyte injury and protection are associated with regulation of COX-2."

    Article Title: Albumin-induced podocyte injury and protection are associated with regulation of COX-2.

    Journal: Kidney international

    doi: 10.1038/ki.2014.196

    SA-associated factors contribute to COX-2 induction in podocytes. Serum-starved podocytes were exposed to 40 mg/ml of BSA, charcoal-treated FA/endotoxin-free BSA, FA/globulin-free BSA, endotoxin-free BSA, HSA and recombinant HSA made in yeast (rHSA). (A) Cells were harvested after 4 h, processed for SDS-PAGE and western blotting and analyzed for COX-2 and GAPDH. (B) Total RNA was extracted and COX-2 mRNA was measured by RT-PCR and normalized to the β-actin mRNA (***P
    Figure Legend Snippet: SA-associated factors contribute to COX-2 induction in podocytes. Serum-starved podocytes were exposed to 40 mg/ml of BSA, charcoal-treated FA/endotoxin-free BSA, FA/globulin-free BSA, endotoxin-free BSA, HSA and recombinant HSA made in yeast (rHSA). (A) Cells were harvested after 4 h, processed for SDS-PAGE and western blotting and analyzed for COX-2 and GAPDH. (B) Total RNA was extracted and COX-2 mRNA was measured by RT-PCR and normalized to the β-actin mRNA (***P

    Techniques Used: Recombinant, SDS Page, Western Blot, Reverse Transcription Polymerase Chain Reaction

    2) Product Images from "Experimental selection of long-term intracellular mycobacteria"

    Article Title: Experimental selection of long-term intracellular mycobacteria

    Journal: Cellular Microbiology

    doi: 10.1111/cmi.12303

    Selected bacteria reside in LC3-positive phagolysosomes.A and B. Long-term infected (LTI) macrophages for at least 2 years and recently infected (RI) with BCG-GFP ancestral strain for 24 h were stained with Lysotracker (A, left panel) or DQ-BSA (B, right panel).C and D. Quantitative analysis of the fluorescence intensity of Lysotracker (C) and DQ-BSA (D) associated with phagosomes in LTI macrophages compared with RI cells. Around 80 phagosomes were quantified in Lysotracker or DQ-BSA stained macrophages from three independent experiments.E and F. Recently infected macrophages (RI) during 24 h (E) and LTI (F) macrophages (2 years old) were subjected to immunofluorescence and endogenous LC3 and LAMP-2 were detected using specific antibodies. Insets depict colocalization with single markers.G and H. Quantitative analysis of the fluorescence intensity of LC3 (G) and LAMP-2 (H) associated with phagosomes of RI and LTI macrophages. Around 130 phagosomes were quantified for LC3 detection and 190 phagosomes for LAMP-2 detection from three independent experiments.Data represent the mean ± S.E.M., (**) P ≤ 0.01, (***) P ≤ 0.001 and (****) P ≤ 0.0001 from two-tailed Student's t -test. Nuclei were stained with Hoechst 33258. Scale bars: 10 μm.
    Figure Legend Snippet: Selected bacteria reside in LC3-positive phagolysosomes.A and B. Long-term infected (LTI) macrophages for at least 2 years and recently infected (RI) with BCG-GFP ancestral strain for 24 h were stained with Lysotracker (A, left panel) or DQ-BSA (B, right panel).C and D. Quantitative analysis of the fluorescence intensity of Lysotracker (C) and DQ-BSA (D) associated with phagosomes in LTI macrophages compared with RI cells. Around 80 phagosomes were quantified in Lysotracker or DQ-BSA stained macrophages from three independent experiments.E and F. Recently infected macrophages (RI) during 24 h (E) and LTI (F) macrophages (2 years old) were subjected to immunofluorescence and endogenous LC3 and LAMP-2 were detected using specific antibodies. Insets depict colocalization with single markers.G and H. Quantitative analysis of the fluorescence intensity of LC3 (G) and LAMP-2 (H) associated with phagosomes of RI and LTI macrophages. Around 130 phagosomes were quantified for LC3 detection and 190 phagosomes for LAMP-2 detection from three independent experiments.Data represent the mean ± S.E.M., (**) P ≤ 0.01, (***) P ≤ 0.001 and (****) P ≤ 0.0001 from two-tailed Student's t -test. Nuclei were stained with Hoechst 33258. Scale bars: 10 μm.

    Techniques Used: Infection, Staining, Fluorescence, Immunofluorescence, Two Tailed Test

    3) Product Images from "The Adenylate Cyclase (CyaA) Toxin from Bordetella pertussis Has No Detectable Phospholipase A (PLA) Activity In Vitro"

    Article Title: The Adenylate Cyclase (CyaA) Toxin from Bordetella pertussis Has No Detectable Phospholipase A (PLA) Activity In Vitro

    Journal: Toxins

    doi: 10.3390/toxins11020111

    The kinetics of fluorescence changes of PED6 incorporated in DOPC:PED6 8:2 LUV (data from Figure S1 ). The time 0 min corresponds to the addition of the following samples: buffer (light blue), urea (dark blue), BSA (green), Crotoxin (black), CyaA IP (orange), and CyaA UBC (red). ( A ) Increase of fluorescence emission intensity at 515 nm. ( B ) Ratio of fluorescence intensity at 515 nm, F 515 , at time t normalized by initial fluorescence at 515 nm. Inset shows same data with an expanded Y -axis (from 0.95 and 1.05). ( C ) Ratio of fluorescence intensity at 515 nm over fluorescence intensity at 575 nm: F 515 /F 575 . ( D ) The F 515 /F 575 values are normalized to the initial F 515 /F 575 value at 0 min (see methods for details).
    Figure Legend Snippet: The kinetics of fluorescence changes of PED6 incorporated in DOPC:PED6 8:2 LUV (data from Figure S1 ). The time 0 min corresponds to the addition of the following samples: buffer (light blue), urea (dark blue), BSA (green), Crotoxin (black), CyaA IP (orange), and CyaA UBC (red). ( A ) Increase of fluorescence emission intensity at 515 nm. ( B ) Ratio of fluorescence intensity at 515 nm, F 515 , at time t normalized by initial fluorescence at 515 nm. Inset shows same data with an expanded Y -axis (from 0.95 and 1.05). ( C ) Ratio of fluorescence intensity at 515 nm over fluorescence intensity at 575 nm: F 515 /F 575 . ( D ) The F 515 /F 575 values are normalized to the initial F 515 /F 575 value at 0 min (see methods for details).

    Techniques Used: Fluorescence

    4) Product Images from "In situ measurement of bovine serum albumin interaction with gold nanospheres"

    Article Title: In situ measurement of bovine serum albumin interaction with gold nanospheres

    Journal: Langmuir

    doi: 10.1021/la3005213

    Autocorrelation curves of AuNPs and AuNPs + BSA [0.75 mM], for three AuNPs samples with mean diameters of 51 nm (A), 70 nm (B) and 93 nm (C). The amplitude of the autocorrelation is normalized to 1 for better comparison of the characteristic diffusion
    Figure Legend Snippet: Autocorrelation curves of AuNPs and AuNPs + BSA [0.75 mM], for three AuNPs samples with mean diameters of 51 nm (A), 70 nm (B) and 93 nm (C). The amplitude of the autocorrelation is normalized to 1 for better comparison of the characteristic diffusion

    Techniques Used: Diffusion-based Assay

    Blip intensity frequency analysis (BIFA) of 51 nm AuNPs before (A) and after (B) binding to BSA [0.75 mM]. The events are counted based on 40 s intensity transients displayed in the insets. (C) Average number of events before (blue) and after (red) binding
    Figure Legend Snippet: Blip intensity frequency analysis (BIFA) of 51 nm AuNPs before (A) and after (B) binding to BSA [0.75 mM]. The events are counted based on 40 s intensity transients displayed in the insets. (C) Average number of events before (blue) and after (red) binding

    Techniques Used: Binding Assay

    (A) Normalized UV/VIS extinction spectra of 56 nm PEG-coated AuNPs before (solid green line) and after (dashed red line) addition of BSA. The inset zooms into the region of λ max . (B) Autocorrelation curves of 56 nm PEG-coated AuNPs before (green)
    Figure Legend Snippet: (A) Normalized UV/VIS extinction spectra of 56 nm PEG-coated AuNPs before (solid green line) and after (dashed red line) addition of BSA. The inset zooms into the region of λ max . (B) Autocorrelation curves of 56 nm PEG-coated AuNPs before (green)

    Techniques Used:

    5) Product Images from "The antibodies against 5-bromo-2′-deoxyuridine specifically recognize trifluridine incorporated into DNA"

    Article Title: The antibodies against 5-bromo-2′-deoxyuridine specifically recognize trifluridine incorporated into DNA

    Journal: Scientific Reports

    doi: 10.1038/srep25286

    Detection of FTD by anti-BrdU antibodies. ( A ) Dot blot analysis of chemically synthesized FTD. BSA-conjugated FTD or non-conjugated (NC) BSA on nitrocellulose membrane were blotted with anti-BrdU antibodies. ( B ) Dot blot analysis of FTD or BrdU incorporated into the genomic DNA of HCT-116 cells. HCT-116 cells were cultured in the presence of 5 μM FTD or BrdU for 4 hours and genomic DNA was purified. Purified DNA was denatured with alkaline solution (0.1 N NaOH), spotted onto Hybond-N + membrane and blotted with anti-BrdU antibodies.
    Figure Legend Snippet: Detection of FTD by anti-BrdU antibodies. ( A ) Dot blot analysis of chemically synthesized FTD. BSA-conjugated FTD or non-conjugated (NC) BSA on nitrocellulose membrane were blotted with anti-BrdU antibodies. ( B ) Dot blot analysis of FTD or BrdU incorporated into the genomic DNA of HCT-116 cells. HCT-116 cells were cultured in the presence of 5 μM FTD or BrdU for 4 hours and genomic DNA was purified. Purified DNA was denatured with alkaline solution (0.1 N NaOH), spotted onto Hybond-N + membrane and blotted with anti-BrdU antibodies.

    Techniques Used: Dot Blot, Synthesized, Cell Culture, Purification

    6) Product Images from "Comparison of the Sensitivities of WaterLOGSY and Saturation Transfer Difference NMR Experiments"

    Article Title: Comparison of the Sensitivities of WaterLOGSY and Saturation Transfer Difference NMR Experiments

    Journal: Journal of biomolecular NMR

    doi: 10.1007/s10858-014-9848-9

    Examples of WaterLOGSY and STD spectra used to compare sensitivities for (a) KET-BSA, (b) TBHQ-HA and (c) CAM-Ribosome 70S. In all cases the relaxation delays, saturation/mixing times, receiver gain and total experimental times were identical.
    Figure Legend Snippet: Examples of WaterLOGSY and STD spectra used to compare sensitivities for (a) KET-BSA, (b) TBHQ-HA and (c) CAM-Ribosome 70S. In all cases the relaxation delays, saturation/mixing times, receiver gain and total experimental times were identical.

    Techniques Used: Chick Chorioallantoic Membrane Assay

    7) Product Images from "Transcript cleavage factors GreA and GreB act as transient catalytic components of RNA polymerase"

    Article Title: Transcript cleavage factors GreA and GreB act as transient catalytic components of RNA polymerase

    Journal: The EMBO Journal

    doi: 10.1093/emboj/cdg610

    Fig. 1. Site-specific GreB–RNAP photocrosslinking. ( A ) Autoradiogram of 8% Tris–glycine SDS–PAGE after UV-irradiation of nine [ 35 S]ASDPC-derivatized GreB-Cys mutants in the presence of RNAP core and BSA as a carrier protein. Numbers on top of the gel indicate the position of the Cys substitution in GreB. GreB-68 is a wt GreB used as a negative control. Positions of free β, β′, BSA and GreB are indicated by arrows. ( B ), shown as ribbons and a schematic representation of their crosslinking targets.
    Figure Legend Snippet: Fig. 1. Site-specific GreB–RNAP photocrosslinking. ( A ) Autoradiogram of 8% Tris–glycine SDS–PAGE after UV-irradiation of nine [ 35 S]ASDPC-derivatized GreB-Cys mutants in the presence of RNAP core and BSA as a carrier protein. Numbers on top of the gel indicate the position of the Cys substitution in GreB. GreB-68 is a wt GreB used as a negative control. Positions of free β, β′, BSA and GreB are indicated by arrows. ( B ), shown as ribbons and a schematic representation of their crosslinking targets.

    Techniques Used: SDS Page, Irradiation, Negative Control

    8) Product Images from "Fabrication of enzyme-degradable and size-controlled protein nanowires using single particle nano-fabrication technique"

    Article Title: Fabrication of enzyme-degradable and size-controlled protein nanowires using single particle nano-fabrication technique

    Journal: Nature Communications

    doi: 10.1038/ncomms4718

    Fabrication of various protein nanowires. AFM images of protein nanowires based on ( a ) BSA, ( b ) OVA, ( c ) avidin.
    Figure Legend Snippet: Fabrication of various protein nanowires. AFM images of protein nanowires based on ( a ) BSA, ( b ) OVA, ( c ) avidin.

    Techniques Used: Avidin-Biotin Assay

    9) Product Images from "Procollagen Triple Helix Assembly: An Unconventional Chaperone-Assisted Folding Paradigm"

    Article Title: Procollagen Triple Helix Assembly: An Unconventional Chaperone-Assisted Folding Paradigm

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0001029

    Procollagen triple helix spontaneously refolds below but not above 34°C. A. Kinetics of 0.1 mg/ml procollagen refolding in PBS after 10 min denaturation of the triple helices at 45°C (monitored by CD as in Fig. 4 ). B and C. Native triple helix refolding in PBS without (B) and with 90 mg/ml BSA (C) after an initial DSC scan from 25 to 50°C at 1°C/min. The fraction of refolded native procollagen (insets) was measured from the area under the DSC thermograms (colored tracings) after overnight equilibration in the DSC instrument at indicated temperatures following the initial denaturation scan (native control). A second scan without the overnight equilibration is shown by the yellow line in C.
    Figure Legend Snippet: Procollagen triple helix spontaneously refolds below but not above 34°C. A. Kinetics of 0.1 mg/ml procollagen refolding in PBS after 10 min denaturation of the triple helices at 45°C (monitored by CD as in Fig. 4 ). B and C. Native triple helix refolding in PBS without (B) and with 90 mg/ml BSA (C) after an initial DSC scan from 25 to 50°C at 1°C/min. The fraction of refolded native procollagen (insets) was measured from the area under the DSC thermograms (colored tracings) after overnight equilibration in the DSC instrument at indicated temperatures following the initial denaturation scan (native control). A second scan without the overnight equilibration is shown by the yellow line in C.

    Techniques Used:

    ER-like molecular crowding with nonspecific proteins does not affect procollagen thermal stability. A and B. Normalized DSC thermograms (A) and apparent T m (B) at 1°C/min heating in PBS without and with 90 mg/ml BSA, 90 mg/ml IgG, or 100 mg/ml lysozyme (the thermograms in A and the corresponding bars in B have the same colors). C. Unfolding kinetics of procollagen at 37.5°C. In PBS with 90 mg/ml BSA, native procollagen fractions (inset, squares) were measured from the area under 1°C/min DSC thermograms of sample aliquots (blue tracings). In PBS without BSA, native procollagen fractions were measured from CD as shown in Fig. 1B .
    Figure Legend Snippet: ER-like molecular crowding with nonspecific proteins does not affect procollagen thermal stability. A and B. Normalized DSC thermograms (A) and apparent T m (B) at 1°C/min heating in PBS without and with 90 mg/ml BSA, 90 mg/ml IgG, or 100 mg/ml lysozyme (the thermograms in A and the corresponding bars in B have the same colors). C. Unfolding kinetics of procollagen at 37.5°C. In PBS with 90 mg/ml BSA, native procollagen fractions (inset, squares) were measured from the area under 1°C/min DSC thermograms of sample aliquots (blue tracings). In PBS without BSA, native procollagen fractions were measured from CD as shown in Fig. 1B .

    Techniques Used:

    10) Product Images from "Hairpin structure within the 3?UTR of DNA polymerase ? mRNA acts as a post-transcriptional regulatory element and interacts with Hax-1"

    Article Title: Hairpin structure within the 3?UTR of DNA polymerase ? mRNA acts as a post-transcriptional regulatory element and interacts with Hax-1

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkm502

    Purified Hax-1 binds to the M2 hairpin structure present in the 3′UTR of the Pol β transcript. UV cross-linking of labeled transcript containing the M2 hairpin structure with purified, recombinant Hax-1. Lanes: 1–9: UV-cross-linked; 1,4,7: with BSA 0.5 μg; 2,5,8: with thioredoxin, 0.5 μg; 3,6,9: with Hax-1 0.5 μg; 1,2,3: H (hairpin); 4,5,6: A (antisense); 7,8,9: Hmut (mutant). The band in lane 3 indicates the binding of recombinant Hax-1 to the hairpin element in the 3′UTR; The band is absent in lane 9—disruption of the hairpin abrogates the binding. Controls with antisense RNA, thioredoxin and BSA do not show any interactions.
    Figure Legend Snippet: Purified Hax-1 binds to the M2 hairpin structure present in the 3′UTR of the Pol β transcript. UV cross-linking of labeled transcript containing the M2 hairpin structure with purified, recombinant Hax-1. Lanes: 1–9: UV-cross-linked; 1,4,7: with BSA 0.5 μg; 2,5,8: with thioredoxin, 0.5 μg; 3,6,9: with Hax-1 0.5 μg; 1,2,3: H (hairpin); 4,5,6: A (antisense); 7,8,9: Hmut (mutant). The band in lane 3 indicates the binding of recombinant Hax-1 to the hairpin element in the 3′UTR; The band is absent in lane 9—disruption of the hairpin abrogates the binding. Controls with antisense RNA, thioredoxin and BSA do not show any interactions.

    Techniques Used: Purification, Labeling, Recombinant, Mutagenesis, Binding Assay

    11) Product Images from "Anti-Inflammatory and Vasoprotective Activity of a Retroviral-Derived Peptide, Homologous to Human Endogenous Retroviruses: Endothelial Cell Effects"

    Article Title: Anti-Inflammatory and Vasoprotective Activity of a Retroviral-Derived Peptide, Homologous to Human Endogenous Retroviruses: Endothelial Cell Effects

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0052693

    MN10021 inhibits dermal vascular leak in a reverse passive Arthus reaction. Groups of 5 Hartley strain guinea pigs (male; ∼250 g) were injected s.c. with 1.0 ml of either saline or the indicated dose of MN10021. Eighteen hours later the flanks of the animals were shaved and both 6.25 and 12.5 µl of rabbit anti-BSA antiserum injected i.d. at each of two sites, one on each flank. The animals were then injected i.v. with 1.0 ml of PBS containing BSA (1.0 mg/ml) and Evans Blue dye (2.0 mg/ml). Six hours later the animals were euthanized by barbiturate overdose, the skin on the back removed, and the diameter of the extravasation of the Evans Blue dye measured for each site on the ventral surface of the skin. The values represent the mean ± SE of the 10 measurements obtained for each of the treatment conditions. ** p
    Figure Legend Snippet: MN10021 inhibits dermal vascular leak in a reverse passive Arthus reaction. Groups of 5 Hartley strain guinea pigs (male; ∼250 g) were injected s.c. with 1.0 ml of either saline or the indicated dose of MN10021. Eighteen hours later the flanks of the animals were shaved and both 6.25 and 12.5 µl of rabbit anti-BSA antiserum injected i.d. at each of two sites, one on each flank. The animals were then injected i.v. with 1.0 ml of PBS containing BSA (1.0 mg/ml) and Evans Blue dye (2.0 mg/ml). Six hours later the animals were euthanized by barbiturate overdose, the skin on the back removed, and the diameter of the extravasation of the Evans Blue dye measured for each site on the ventral surface of the skin. The values represent the mean ± SE of the 10 measurements obtained for each of the treatment conditions. ** p

    Techniques Used: Injection

    Binding of [ 125 I] – MN20054 to Fibroblasts. HUVEC and WI-38 fibroblasts (CCL-75™; ATCC) were grown to confluence in 12-well tissue culture plates. To each of triplicate wells was added 20 pmol of [ 125 I]-MN20054 in the presence of either buffer alone, a 100-fold molar excess of MN20054, or a 100-fold molar excess of MN10021. The plates were incubated for 60 min at 37°C with gentle rocking, the wells aspirated and quickly washed 3X with 2.0 ml each of ice-cold PBS/BSA and 1.0 ml of 1.0 N NaOH added to each well to solubilize the cells. Nine-tenths ml of solubilized cells was then removed from each well and cell-associated radioactivity determined by gamma scintillation spectrophotometry.
    Figure Legend Snippet: Binding of [ 125 I] – MN20054 to Fibroblasts. HUVEC and WI-38 fibroblasts (CCL-75™; ATCC) were grown to confluence in 12-well tissue culture plates. To each of triplicate wells was added 20 pmol of [ 125 I]-MN20054 in the presence of either buffer alone, a 100-fold molar excess of MN20054, or a 100-fold molar excess of MN10021. The plates were incubated for 60 min at 37°C with gentle rocking, the wells aspirated and quickly washed 3X with 2.0 ml each of ice-cold PBS/BSA and 1.0 ml of 1.0 N NaOH added to each well to solubilize the cells. Nine-tenths ml of solubilized cells was then removed from each well and cell-associated radioactivity determined by gamma scintillation spectrophotometry.

    Techniques Used: Binding Assay, Incubation, Radioactivity, Spectrophotometry

    Monocyte and endothelial cell binding of MN10021 and MN20054. A. Human blood monocytes, isolated as described in Methods, were adjusted to 5×10 6 /ml in PBS (0.5% BSA, pH 7.5). Two-tenths ml of cell suspension was added to each of triplicate 12×75 mm polypropylene tubes and the tubes were incubated for 60 min at 37°C with 20 pmol of [ 125 I]-MN20054 in the presence of either buffer alone, a 100-fold molar excess of MN20054, a 100-fold molar excess of MN10021, or a 100-fold molar excess of MN20050. The contents of each tube were then layered onto 0.8 ml of 10% sucrose in Eppendorf tubes, centrifuged, and the cell-associated radioactivity determined by gamma scintillation spectrophotometry of the cell pellet. B. Binding/uptake of [ 125 I]-MN20054 by human monocytes was performed as described for panel A with the following exceptions: (1) half the cells were pretreated for 30 min at 37°C with cytochalasin B to inhibit endocytosis and (2) competition was performed only with a 100-fold molar excess of MN10021. C. HUVEC were grown to confluence in 12-well tissue culture plates. To each of triplicate wells was added 20 pmol of [ 125 I]-MN20054 in the presence of either buffer alone, a 100-fold molar excess of MN20054, a 100-fold molar excess of MN10021, or a 100-fold molar excess of MN20050. The plates were incubated for 60 min at 37°C with gentle rocking, the wells aspirated and quickly washed 3X with 2.0 ml each of ice-cold PBS/BSA and 1.0 ml of 1.0 N NaOH added to each well to solubilize the cells. Nine-tenths ml of solubilized cells was then removed from each well and cell-associated radioactivity determined by gamma scintillation spectrophotometry.
    Figure Legend Snippet: Monocyte and endothelial cell binding of MN10021 and MN20054. A. Human blood monocytes, isolated as described in Methods, were adjusted to 5×10 6 /ml in PBS (0.5% BSA, pH 7.5). Two-tenths ml of cell suspension was added to each of triplicate 12×75 mm polypropylene tubes and the tubes were incubated for 60 min at 37°C with 20 pmol of [ 125 I]-MN20054 in the presence of either buffer alone, a 100-fold molar excess of MN20054, a 100-fold molar excess of MN10021, or a 100-fold molar excess of MN20050. The contents of each tube were then layered onto 0.8 ml of 10% sucrose in Eppendorf tubes, centrifuged, and the cell-associated radioactivity determined by gamma scintillation spectrophotometry of the cell pellet. B. Binding/uptake of [ 125 I]-MN20054 by human monocytes was performed as described for panel A with the following exceptions: (1) half the cells were pretreated for 30 min at 37°C with cytochalasin B to inhibit endocytosis and (2) competition was performed only with a 100-fold molar excess of MN10021. C. HUVEC were grown to confluence in 12-well tissue culture plates. To each of triplicate wells was added 20 pmol of [ 125 I]-MN20054 in the presence of either buffer alone, a 100-fold molar excess of MN20054, a 100-fold molar excess of MN10021, or a 100-fold molar excess of MN20050. The plates were incubated for 60 min at 37°C with gentle rocking, the wells aspirated and quickly washed 3X with 2.0 ml each of ice-cold PBS/BSA and 1.0 ml of 1.0 N NaOH added to each well to solubilize the cells. Nine-tenths ml of solubilized cells was then removed from each well and cell-associated radioactivity determined by gamma scintillation spectrophotometry.

    Techniques Used: Binding Assay, Isolation, Incubation, Radioactivity, Spectrophotometry

    12) Product Images from "Role of the receptor for advanced glycation end products in hepatic fibrosis"

    Article Title: Role of the receptor for advanced glycation end products in hepatic fibrosis

    Journal: World Journal of Gastroenterology : WJG

    doi: 10.3748/wjg.15.5789

    AGE-BSA does not activate p44/42 (A) and p38 (B) MAPK signaling in hepatic stellate cells. Kinase phosphorylation was determined in CFSC-2G and HSC-T6 HSC using quantitative western blotting with phosphospecific antibodies relative to total kinase antibodies
    Figure Legend Snippet: AGE-BSA does not activate p44/42 (A) and p38 (B) MAPK signaling in hepatic stellate cells. Kinase phosphorylation was determined in CFSC-2G and HSC-T6 HSC using quantitative western blotting with phosphospecific antibodies relative to total kinase antibodies

    Techniques Used: Western Blot

    AGE do not stimulate DNA synthesis in hepatic stellate cells. DNA synthesis as a surrogate of proliferation was determined in CFSC-2G (A) and HSC-T6 (B) HSC after a 24 h incubation with increasing concentrations of FCS, AGE-BSA or CML-BSA in 0% FCS. Data
    Figure Legend Snippet: AGE do not stimulate DNA synthesis in hepatic stellate cells. DNA synthesis as a surrogate of proliferation was determined in CFSC-2G (A) and HSC-T6 (B) HSC after a 24 h incubation with increasing concentrations of FCS, AGE-BSA or CML-BSA in 0% FCS. Data

    Techniques Used: DNA Synthesis, Incubation

    13) Product Images from "First Total Synthesis of a Naturally Occurring Iodinated 5?-Deoxyxylofuranosyl Marine Nucleoside"

    Article Title: First Total Synthesis of a Naturally Occurring Iodinated 5?-Deoxyxylofuranosyl Marine Nucleoside

    Journal: Marine Drugs

    doi: 10.3390/md10040881

    Total synthesis of pyrrolo[2,3- d ]pyrimidine nucleoside 1 . Reagents and conditions: ( a ) (a1) Conc. H 2 SO 4 , acetone, 0 °C, 3 h; (a2) 5% Na 2 CO 3 aq., 30 min, 87%; ( b ) TsCl (1.1 eq.), THF, Et 3 N, 0 °C, overnight, 92%; ( c ) LiAlH 4 (0.5 eq.), THF, reflux, 6 h, 95%; ( d ) BzCl (1.1 eq), CH 2 Cl 2 , Et 3 N, 2 h, 98%; ( e ) Conc. H 2 SO 4 , Ac 2 O, 26 h, 78%; ( f ) POCl 3 (excess), reflux, 3 h, 98%; ( g ) NIS (1.01 eq.), DMF, 0 °C, 2 h, 98%; ( h ) 4 (1 eq.), BSA (1.2 eq.), TMSOTf (1 eq.), CH 3 CN, rt-80 °C, 12 h 56%; ( i ) Sat. NH 3 in MeOH (excess), 130 °C, 12 h, 82%.
    Figure Legend Snippet: Total synthesis of pyrrolo[2,3- d ]pyrimidine nucleoside 1 . Reagents and conditions: ( a ) (a1) Conc. H 2 SO 4 , acetone, 0 °C, 3 h; (a2) 5% Na 2 CO 3 aq., 30 min, 87%; ( b ) TsCl (1.1 eq.), THF, Et 3 N, 0 °C, overnight, 92%; ( c ) LiAlH 4 (0.5 eq.), THF, reflux, 6 h, 95%; ( d ) BzCl (1.1 eq), CH 2 Cl 2 , Et 3 N, 2 h, 98%; ( e ) Conc. H 2 SO 4 , Ac 2 O, 26 h, 78%; ( f ) POCl 3 (excess), reflux, 3 h, 98%; ( g ) NIS (1.01 eq.), DMF, 0 °C, 2 h, 98%; ( h ) 4 (1 eq.), BSA (1.2 eq.), TMSOTf (1 eq.), CH 3 CN, rt-80 °C, 12 h 56%; ( i ) Sat. NH 3 in MeOH (excess), 130 °C, 12 h, 82%.

    Techniques Used: Reflux

    14) Product Images from "Rational Design of “Three-in-One” Ratiometric Nanoprobes: Protein-Caged Dityrosine, CdS Quantum Dots, and Gold Nanoclusters"

    Article Title: Rational Design of “Three-in-One” Ratiometric Nanoprobes: Protein-Caged Dityrosine, CdS Quantum Dots, and Gold Nanoclusters

    Journal: ACS Omega

    doi: 10.1021/acsomega.0c00711

    (A) Emission spectra of the CdSQDs/AuNCs@diTyr-BSA nanohybrids in the presence of various concentrations of GSH. Inset: Corresponding UV LED light photographs (340 nm) and concentration along the direction of the arrow: 0, 1, 2, 5, 10, 20, 40, 60, 80, 100, 200, 400, 600, 800, 1000, and 1200 μM. (B) Ratio values (FL green /FL red ) of the CdSQDs/AuNCs@diTyr-BSA nanohybrids as a function of GSH concentration. (C) Ratio values (FL green /FL red ) of the CdSQDs/AuNCs@diTyr-BSA nanohybrids in the presence of various underlying interferences at the same concentration (1 mM). (D) Relative viability of MCF-7 cells treated with the indicated amount of CdSQDs/AuNCs@diTyr-BSA nanohybrids. (E) FL green /FL red values in MCF-7 cells obtained on the basis of (F) fluorescence microscope images of MCF-7 cells treated with the CdSQDs/AuNCs@diTyr-BSA nanohybrids without (up panel) and with 0.1 mM BSO treatment.
    Figure Legend Snippet: (A) Emission spectra of the CdSQDs/AuNCs@diTyr-BSA nanohybrids in the presence of various concentrations of GSH. Inset: Corresponding UV LED light photographs (340 nm) and concentration along the direction of the arrow: 0, 1, 2, 5, 10, 20, 40, 60, 80, 100, 200, 400, 600, 800, 1000, and 1200 μM. (B) Ratio values (FL green /FL red ) of the CdSQDs/AuNCs@diTyr-BSA nanohybrids as a function of GSH concentration. (C) Ratio values (FL green /FL red ) of the CdSQDs/AuNCs@diTyr-BSA nanohybrids in the presence of various underlying interferences at the same concentration (1 mM). (D) Relative viability of MCF-7 cells treated with the indicated amount of CdSQDs/AuNCs@diTyr-BSA nanohybrids. (E) FL green /FL red values in MCF-7 cells obtained on the basis of (F) fluorescence microscope images of MCF-7 cells treated with the CdSQDs/AuNCs@diTyr-BSA nanohybrids without (up panel) and with 0.1 mM BSO treatment.

    Techniques Used: Concentration Assay, Fluorescence, Microscopy

    15) Product Images from "Spectral-domain optical coherence phase microscopy for label-free multiplexed protein microarray assay"

    Article Title: Spectral-domain optical coherence phase microscopy for label-free multiplexed protein microarray assay

    Journal: Biosensors & bioelectronics

    doi: 10.1016/j.bios.2009.06.028

    Calculated kinetic association and dissociation parameters for BSA and HSA spots with αHSA. (a) Sensorgrams for each spot. (b) and (c) present k a and k d averaged over 5 spots for each species. The error bar denotes standard deviation.
    Figure Legend Snippet: Calculated kinetic association and dissociation parameters for BSA and HSA spots with αHSA. (a) Sensorgrams for each spot. (b) and (c) present k a and k d averaged over 5 spots for each species. The error bar denotes standard deviation.

    Techniques Used: Standard Deviation

    16) Product Images from "Pharmacological inhibition of adipose triglyceride lipase corrects high-fat diet-induced insulin resistance and hepatosteatosis in mice"

    Article Title: Pharmacological inhibition of adipose triglyceride lipase corrects high-fat diet-induced insulin resistance and hepatosteatosis in mice

    Journal: Nature Communications

    doi: 10.1038/ncomms14859

    Atglistatin transiently inhibits lipolysis and protects from HFD-induced obesity. Six weeks old male C57Bl6J mice were fed a HFD (45 kJ% fat; 22.1 kJ g −1 ) for 50 days. Thereafter, mice were fasted for 7 h and then re-fed a HFD with or without Atglistatin (2 mmol kg −1 diet) for 2 h followed by a second, subsequent fasting period of 8 h. ( a ) Plasma FA levels were determined in the 2 h re-fed and 8 h fasted state ( n =5). ( b ) FA release from gonadal adipose tissue explants of re-fed and ( c ) 8 h-fasted mice ( n =5 per group). ( d , e ) Atglistatin does not inhibit human adipocyte lipolysis. ( d ) TG hydrolase activity was assessed in COS-7 lysates of cells overexpressing human and murine ATGL and CGI-58, respectively, in the presence and absence of the indicated concentrations of Atglistatin. ( e ) SGBS and 3T3-L1 preadipocytes were differentiated to adipocytes. Then, cells were preincubated with the indicated concentrations of Atglistatin for 2 h. Thereafter, the medium was replaced by DMEM containing 2% BSA, 10 μM Forskolin and the indicated concentrations of Atglistatin for 1 h. The release of FA in the medium was determined and calculated per mg cell protein. ( f – k ) Mice were fed a HFD for 50 days, followed by HFD-feeding in the presence or absence of Atglistatin for another 50 days. ( f ) Body weight, and ( g ) fat and lean mass development ( n =7 per group). Adipose tissue depots, inguinal (i)WAT, gonadal (g)WAT, and interscapular (i)BAT were analysed for their ( h ) weight and ( i ) adipocyte size. ( j , k ) mRNA expression of IL-6 and macrophage markers F4/80 and Cd11c was assessed in gWAT and iBAT of re-fed mice ( n =5 per group). Data represent mean±s.d. Statistical significance was determined by Student's two-tailed t -test. For analysis of multiple measurements, we performed one-way analysis of variance (ANOVA) followed by Bonferroni post-hoc test; * P
    Figure Legend Snippet: Atglistatin transiently inhibits lipolysis and protects from HFD-induced obesity. Six weeks old male C57Bl6J mice were fed a HFD (45 kJ% fat; 22.1 kJ g −1 ) for 50 days. Thereafter, mice were fasted for 7 h and then re-fed a HFD with or without Atglistatin (2 mmol kg −1 diet) for 2 h followed by a second, subsequent fasting period of 8 h. ( a ) Plasma FA levels were determined in the 2 h re-fed and 8 h fasted state ( n =5). ( b ) FA release from gonadal adipose tissue explants of re-fed and ( c ) 8 h-fasted mice ( n =5 per group). ( d , e ) Atglistatin does not inhibit human adipocyte lipolysis. ( d ) TG hydrolase activity was assessed in COS-7 lysates of cells overexpressing human and murine ATGL and CGI-58, respectively, in the presence and absence of the indicated concentrations of Atglistatin. ( e ) SGBS and 3T3-L1 preadipocytes were differentiated to adipocytes. Then, cells were preincubated with the indicated concentrations of Atglistatin for 2 h. Thereafter, the medium was replaced by DMEM containing 2% BSA, 10 μM Forskolin and the indicated concentrations of Atglistatin for 1 h. The release of FA in the medium was determined and calculated per mg cell protein. ( f – k ) Mice were fed a HFD for 50 days, followed by HFD-feeding in the presence or absence of Atglistatin for another 50 days. ( f ) Body weight, and ( g ) fat and lean mass development ( n =7 per group). Adipose tissue depots, inguinal (i)WAT, gonadal (g)WAT, and interscapular (i)BAT were analysed for their ( h ) weight and ( i ) adipocyte size. ( j , k ) mRNA expression of IL-6 and macrophage markers F4/80 and Cd11c was assessed in gWAT and iBAT of re-fed mice ( n =5 per group). Data represent mean±s.d. Statistical significance was determined by Student's two-tailed t -test. For analysis of multiple measurements, we performed one-way analysis of variance (ANOVA) followed by Bonferroni post-hoc test; * P

    Techniques Used: Mouse Assay, Activity Assay, Expressing, Two Tailed Test

    17) Product Images from "Enzyme-Linked Immunosorbent Assay (ELISA) and Blocking with Bovine Serum Albumin (BSA) - Not all BSAs are alike"

    Article Title: Enzyme-Linked Immunosorbent Assay (ELISA) and Blocking with Bovine Serum Albumin (BSA) - Not all BSAs are alike

    Journal: Journal of immunological methods

    doi: 10.1016/j.jim.2012.06.009

    Specific vs. non-specific binding of VCP to C3b or various BSA preparations
    Figure Legend Snippet: Specific vs. non-specific binding of VCP to C3b or various BSA preparations

    Techniques Used: Binding Assay

    18) Product Images from "High affinity nanobodies against human epidermal growth factor receptor selected on cells by E. coli display"

    Article Title: High affinity nanobodies against human epidermal growth factor receptor selected on cells by E. coli display

    Journal: mAbs

    doi: 10.1080/19420862.2016.1216742

    Binding of E. coli bacteria displaying selected VHH clones to eEGFR-Fc. Fluorescent flow cytometry analysis of induced E. coli EcM1 cells displaying the indicated Nb (VEGFR1-6) incubated with biotinylated eEGFR-Fc (red) or BSA (black) and secondary Streptavidin-PE.
    Figure Legend Snippet: Binding of E. coli bacteria displaying selected VHH clones to eEGFR-Fc. Fluorescent flow cytometry analysis of induced E. coli EcM1 cells displaying the indicated Nb (VEGFR1-6) incubated with biotinylated eEGFR-Fc (red) or BSA (black) and secondary Streptavidin-PE.

    Techniques Used: Binding Assay, Clone Assay, Flow Cytometry, Cytometry, Incubation

    Binding activity of the selected Nbs to eEGFR-Fc determined by ELISA. Binding curves determined by ELISA using the purified anti-EGFR Nbs with C-terminal 6xHis and myc tags. The ELISA was developed with anti-myc-tag mAb and anti-mouse-POD. ELISA signals against a control antigen (BSA) were subtracted from the represented values. The plot shows the OD values at 490 nm with standard error from 2 independent experiments with duplicates using the purified Nbs at the indicated concentrations (0.05–1000 nM).
    Figure Legend Snippet: Binding activity of the selected Nbs to eEGFR-Fc determined by ELISA. Binding curves determined by ELISA using the purified anti-EGFR Nbs with C-terminal 6xHis and myc tags. The ELISA was developed with anti-myc-tag mAb and anti-mouse-POD. ELISA signals against a control antigen (BSA) were subtracted from the represented values. The plot shows the OD values at 490 nm with standard error from 2 independent experiments with duplicates using the purified Nbs at the indicated concentrations (0.05–1000 nM).

    Techniques Used: Binding Assay, Activity Assay, Enzyme-linked Immunosorbent Assay, Purification

    Competition of EGF and the selected Nbs for binding to eEGFR-Fc determined by flow cytometry analysis of E. coli bacteria. Flow cytometry analysis of induced E. coli EcM1 bacteria displaying the indicated Nb (VEGFR1-6) and incubated with 50 nM of biotinylated BSA (black) or eEGFR-Fc in the absence (red) or presence (blue) of 25 µM EGF. Overlay of histograms showing the fluorescence intensity of bacteria after staining with secondary Streptavidin-PE.
    Figure Legend Snippet: Competition of EGF and the selected Nbs for binding to eEGFR-Fc determined by flow cytometry analysis of E. coli bacteria. Flow cytometry analysis of induced E. coli EcM1 bacteria displaying the indicated Nb (VEGFR1-6) and incubated with 50 nM of biotinylated BSA (black) or eEGFR-Fc in the absence (red) or presence (blue) of 25 µM EGF. Overlay of histograms showing the fluorescence intensity of bacteria after staining with secondary Streptavidin-PE.

    Techniques Used: Binding Assay, Flow Cytometry, Cytometry, Incubation, Fluorescence, Staining

    19) Product Images from "Integrin αvβ3 on human endothelial cells binds von Willebrand factor strings under fluid shear stress"

    Article Title: Integrin αvβ3 on human endothelial cells binds von Willebrand factor strings under fluid shear stress

    Journal: Blood

    doi: 10.1182/blood-2008-05-158584

    P-selectin and VWF string formation . (A) Wells in an enzyme-linked immunosorbent assay plate were coated with either 20 μg/mL purified P-selectin (R D Systems) or 1% BSA. U937 cells, which express P-selectin ligand PSGL-1, were resuspended
    Figure Legend Snippet: P-selectin and VWF string formation . (A) Wells in an enzyme-linked immunosorbent assay plate were coated with either 20 μg/mL purified P-selectin (R D Systems) or 1% BSA. U937 cells, which express P-selectin ligand PSGL-1, were resuspended

    Techniques Used: Enzyme-linked Immunosorbent Assay, Purification

    20) Product Images from "Resolubilization of Protein from Water-Insoluble Phlorotannin–Protein Complexes upon Acidification"

    Article Title: Resolubilization of Protein from Water-Insoluble Phlorotannin–Protein Complexes upon Acidification

    Journal: Journal of Agricultural and Food Chemistry

    doi: 10.1021/acs.jafc.7b03779

    Fluorescence emission spectra (λ ex = 280 nm) of (A) β-casein and (B) BSA quenched by an increasing tannin concentration. Stern–Volmer plots (λ em = 350 nm) for binding of PhT (0–25 μM assuming an average MW = 2000 Da) (●) and PGG (0–50 μM) (□) to (C) β-casein and (D) BSA at pH 8.0.
    Figure Legend Snippet: Fluorescence emission spectra (λ ex = 280 nm) of (A) β-casein and (B) BSA quenched by an increasing tannin concentration. Stern–Volmer plots (λ em = 350 nm) for binding of PhT (0–25 μM assuming an average MW = 2000 Da) (●) and PGG (0–50 μM) (□) to (C) β-casein and (D) BSA at pH 8.0.

    Techniques Used: Fluorescence, Concentration Assay, Binding Assay

    Proportions (%) of (A and C) precipitated protein and (B and D) precipitated PhT (×) after precipitation of protein (□) or mixtures of PhT/protein (○) for (A and B) β-casein and (C and D) BSA as a function of pH. (- - -) pI of the proteins.
    Figure Legend Snippet: Proportions (%) of (A and C) precipitated protein and (B and D) precipitated PhT (×) after precipitation of protein (□) or mixtures of PhT/protein (○) for (A and B) β-casein and (C and D) BSA as a function of pH. (- - -) pI of the proteins.

    Techniques Used:

    (A) Fluorescence spectra of BSA (λ ex = 280 nm) at pH 8 (−), pH 6.0 (- - -), and pH 3.0 (···). Stern–Volmer plots (λ em = 350 nm) of (B) 5 μM β-casein and (C) 5 μM BSA quenched by an increasing PhT concentration (0–25 μM, assuming an average MW = 2000 Da) at pH 8 (●), pH 6 (□), and pH 3 (×).
    Figure Legend Snippet: (A) Fluorescence spectra of BSA (λ ex = 280 nm) at pH 8 (−), pH 6.0 (- - -), and pH 3.0 (···). Stern–Volmer plots (λ em = 350 nm) of (B) 5 μM β-casein and (C) 5 μM BSA quenched by an increasing PhT concentration (0–25 μM, assuming an average MW = 2000 Da) at pH 8 (●), pH 6 (□), and pH 3 (×).

    Techniques Used: Fluorescence, Concentration Assay

    Protein (%) remaining in precipitation after resolubilization from precipitated β-casein (□) from PhT/protein combinations (○) and PhT/protein/PEG combinations (◇) for (A) β-casein and (B) BSA as a function of pH. In absence of tannins, BSA did not precipitate.
    Figure Legend Snippet: Protein (%) remaining in precipitation after resolubilization from precipitated β-casein (□) from PhT/protein combinations (○) and PhT/protein/PEG combinations (◇) for (A) β-casein and (B) BSA as a function of pH. In absence of tannins, BSA did not precipitate.

    Techniques Used:

    21) Product Images from "Lsa21, a novel leptospiral protein binding adhesive matrix molecules and present during human infection"

    Article Title: Lsa21, a novel leptospiral protein binding adhesive matrix molecules and present during human infection

    Journal: BMC Microbiology

    doi: 10.1186/1471-2180-8-70

    Binding of Lsa21 recombinant protein to ECM components . Wells were coated with 1 μg of laminin, collagen Type I, collagen Type IV, cellular fibronectin, plasma fibronectin, and the control proteins BSA and fetuin. Recombinant Lsa21 and Loa22 proteins attachment to those ECM macromolecules was assessed by an ELISA-based assay. One microgram of recombinant protein was added per well. Optical densities were taken at 492 nm. Data represent the mean ± standard error of three independent experiments. For statistical analysis, the attachment of Lsa21 to ECM macromolecules was compared to its binding to BSA by the Student's two tailed t - test (*, P
    Figure Legend Snippet: Binding of Lsa21 recombinant protein to ECM components . Wells were coated with 1 μg of laminin, collagen Type I, collagen Type IV, cellular fibronectin, plasma fibronectin, and the control proteins BSA and fetuin. Recombinant Lsa21 and Loa22 proteins attachment to those ECM macromolecules was assessed by an ELISA-based assay. One microgram of recombinant protein was added per well. Optical densities were taken at 492 nm. Data represent the mean ± standard error of three independent experiments. For statistical analysis, the attachment of Lsa21 to ECM macromolecules was compared to its binding to BSA by the Student's two tailed t - test (*, P

    Techniques Used: Binding Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Two Tailed Test

    (A) Lsa21 binds to ECM components in a dose-dependent manner . Binding of Lsa21 to laminin, collagen IV and plasma fibronectin, as a function of protein concentration (0 to 2,000 nM). Each point represents the mean absorbance value at 492 nm ± the standard deviation of three independent experiments. BSA was included as a negative control. (B) Sugar moiety contribution to the laminin-Lsa21 interaction . Immobilized laminin was treated with sodium metaperiodate (5 to 100 mM) for 15 min at 4°C in the dark. Each point represents the mean absorbance value at 492 nm ± the standard deviation of three independent experiments. Reduction in attachment compared to the level of attachment to untreated laminin was statistically significant: in each case (*), the P value, as measured by the Student two-tailed t -test, was ≤ 0.004.
    Figure Legend Snippet: (A) Lsa21 binds to ECM components in a dose-dependent manner . Binding of Lsa21 to laminin, collagen IV and plasma fibronectin, as a function of protein concentration (0 to 2,000 nM). Each point represents the mean absorbance value at 492 nm ± the standard deviation of three independent experiments. BSA was included as a negative control. (B) Sugar moiety contribution to the laminin-Lsa21 interaction . Immobilized laminin was treated with sodium metaperiodate (5 to 100 mM) for 15 min at 4°C in the dark. Each point represents the mean absorbance value at 492 nm ± the standard deviation of three independent experiments. Reduction in attachment compared to the level of attachment to untreated laminin was statistically significant: in each case (*), the P value, as measured by the Student two-tailed t -test, was ≤ 0.004.

    Techniques Used: Binding Assay, Protein Concentration, Standard Deviation, Negative Control, Two Tailed Test

    22) Product Images from "Cerebrovascular amyloid Angiopathy in bioengineered vessels is reduced by high-density lipoprotein particles enriched in Apolipoprotein E"

    Article Title: Cerebrovascular amyloid Angiopathy in bioengineered vessels is reduced by high-density lipoprotein particles enriched in Apolipoprotein E

    Journal: Molecular Neurodegeneration

    doi: 10.1186/s13024-020-00366-8

    HDL binds to both col lagen-I and to Aβ42 to form a complex and increases Aβ transport through the bioengineered vessel wall primarily by HDL-apoE particles. a SMC were grown for 3 d on 2D coverslips before incubating with 0.5 μM FITC-Aβ42 (depicted as yellow) and 200 μg/ml Alexa-633 labeled HDL (depicted as cyan). After 24 h, cells were fixed and stained for collagen-I (magenta) before imaging by confocal microscopy. White arrows show co-localization of Aβ42, HDL and collagen-I. b Black 96 well-plates were coated either with 50 μg/mL rat-tail collagen-I or 10% BSA as protein control for 24 h before incubating with a solution of 1 μM of FITC-Aβ42 with or without 200 μg/mL of HDL. After 24 h and extensive PBS washes, fluorescence representing bound Aβ42 was measured at 520 nm, excitation 490 nm. c 1 μM Aβ42 were incubated either with 200 μg/mL of HDL or BSA for 24 h at 37 °C before gel-filtration chromatography separation. Fractions were dot blotted and immunodetected for Aβ (6E10) or HDL (apoA-II). The intensity of each fraction on the dot blot was quantified, normalized between 0 and 100% and graphed on the left panel. The right panel shows a representative dot blot used for quantification. d 1 μM Aβ42 was injected in the tissue chamber of bioengineered vessels and 200 μg/mL of HDL was circulated through the lumen. After 24 h, tissues were homogenized in TBS to collect soluble Aβ, which was quantified in the tissue ( c ) or circulating media ( d ) by ELISA. f 1 μM Aβ42 was injected in the tissue chamber of bioengineered vessels and 200 μg/mL of HDL was circulated through the lumen. After 24 h vessels were lysed in RIPA and protein distribution was analysed using non-denaturing native blot probed for collagen-I, apoE, apoA-II, apoA-I and Aβ (6E10). g HDL was enriched for or depleted of apoE using an apoE immunoaffinity column. Bioengineered vessels were then treated with either fraction (200 μg/mL) in the presence of 1 μM Aβ42 as above before measuring Aβ deposition in the RIPA-soluble fraction. h RIPA lysates from engineered vessels were then analysed using non-denaturing native blot probed for collagen-I and apoE. i Black 96-well plates were coated either with 50 μg/mL rat-tail collagen-I or 10% BSA for 24 h before incubating with a solution of 1 μM of FITC-Aβ42 with or without 200 μg/ml of total HDL, apoE-depleted HDL or apoE-enriched HDL. After 24 h and extensive PBS washes, fluorescence was measured at 520 nm, excitation 490 nm. FPLC data and Western blots are representative of at least 3 individual experiments. Points in graphed data represent individual bioengineered vessels, bars represent mean, error bars represent ±SEM and analysed by Student’s t-test or one way ANOVA * P
    Figure Legend Snippet: HDL binds to both col lagen-I and to Aβ42 to form a complex and increases Aβ transport through the bioengineered vessel wall primarily by HDL-apoE particles. a SMC were grown for 3 d on 2D coverslips before incubating with 0.5 μM FITC-Aβ42 (depicted as yellow) and 200 μg/ml Alexa-633 labeled HDL (depicted as cyan). After 24 h, cells were fixed and stained for collagen-I (magenta) before imaging by confocal microscopy. White arrows show co-localization of Aβ42, HDL and collagen-I. b Black 96 well-plates were coated either with 50 μg/mL rat-tail collagen-I or 10% BSA as protein control for 24 h before incubating with a solution of 1 μM of FITC-Aβ42 with or without 200 μg/mL of HDL. After 24 h and extensive PBS washes, fluorescence representing bound Aβ42 was measured at 520 nm, excitation 490 nm. c 1 μM Aβ42 were incubated either with 200 μg/mL of HDL or BSA for 24 h at 37 °C before gel-filtration chromatography separation. Fractions were dot blotted and immunodetected for Aβ (6E10) or HDL (apoA-II). The intensity of each fraction on the dot blot was quantified, normalized between 0 and 100% and graphed on the left panel. The right panel shows a representative dot blot used for quantification. d 1 μM Aβ42 was injected in the tissue chamber of bioengineered vessels and 200 μg/mL of HDL was circulated through the lumen. After 24 h, tissues were homogenized in TBS to collect soluble Aβ, which was quantified in the tissue ( c ) or circulating media ( d ) by ELISA. f 1 μM Aβ42 was injected in the tissue chamber of bioengineered vessels and 200 μg/mL of HDL was circulated through the lumen. After 24 h vessels were lysed in RIPA and protein distribution was analysed using non-denaturing native blot probed for collagen-I, apoE, apoA-II, apoA-I and Aβ (6E10). g HDL was enriched for or depleted of apoE using an apoE immunoaffinity column. Bioengineered vessels were then treated with either fraction (200 μg/mL) in the presence of 1 μM Aβ42 as above before measuring Aβ deposition in the RIPA-soluble fraction. h RIPA lysates from engineered vessels were then analysed using non-denaturing native blot probed for collagen-I and apoE. i Black 96-well plates were coated either with 50 μg/mL rat-tail collagen-I or 10% BSA for 24 h before incubating with a solution of 1 μM of FITC-Aβ42 with or without 200 μg/ml of total HDL, apoE-depleted HDL or apoE-enriched HDL. After 24 h and extensive PBS washes, fluorescence was measured at 520 nm, excitation 490 nm. FPLC data and Western blots are representative of at least 3 individual experiments. Points in graphed data represent individual bioengineered vessels, bars represent mean, error bars represent ±SEM and analysed by Student’s t-test or one way ANOVA * P

    Techniques Used: Labeling, Staining, Imaging, Confocal Microscopy, Fluorescence, Incubation, Filtration, Chromatography, Dot Blot, Injection, Enzyme-linked Immunosorbent Assay, Fast Protein Liquid Chromatography, Western Blot

    23) Product Images from "Direct Oxidation and Covalent Binding of Isoniazid to Rodent Liver and Human Hepatic Microsomes: Humans Are More Like Mice than Rats"

    Article Title: Direct Oxidation and Covalent Binding of Isoniazid to Rodent Liver and Human Hepatic Microsomes: Humans Are More Like Mice than Rats

    Journal: Chemical Research in Toxicology

    doi: 10.1021/tx300341r

    Specificity of the anti-INH antibody. (A) Antibody specificity was tested by ELISA. The plate was either coated with BSA modified with INA (BSA-INA) or BSA alone. Preimmune serum (S Pre ) or serum after immunization with Blue Carrier Protein modified with INA (S Aft ) diluted at 1:100,000 was used as the primary antibody. In the third and fifth column, the primary antibody was preincubated with INH or NAL, respectively, at a concentration of 200 μM for 30 min at room temperature. (B) Antibody was tested for cross-reactivity with binding due to AcHz. Female C57BL/6 mice were treated with either INH or AcHz (Fisher Scientific) by gavage for 7 days at 50 mg/kg/day ( n = 2 for each group). There was no binding to hepatic proteins from untreated control and AcHz-treated mice, whereas hepatic proteins from INH-treated mice showed a large number of bands modified with INH. (C) Antiserum was tested for specificity on Western blots by preincubation with INH at 200 μM or 2 mM for 1 h at 4 °C, which prevented binding to the INH-modified hepatic proteins. (D) Binding of the anti-INH serum to INH-modified liver proteins on Western blots was compared to that of the preimmune serum from the same animal.
    Figure Legend Snippet: Specificity of the anti-INH antibody. (A) Antibody specificity was tested by ELISA. The plate was either coated with BSA modified with INA (BSA-INA) or BSA alone. Preimmune serum (S Pre ) or serum after immunization with Blue Carrier Protein modified with INA (S Aft ) diluted at 1:100,000 was used as the primary antibody. In the third and fifth column, the primary antibody was preincubated with INH or NAL, respectively, at a concentration of 200 μM for 30 min at room temperature. (B) Antibody was tested for cross-reactivity with binding due to AcHz. Female C57BL/6 mice were treated with either INH or AcHz (Fisher Scientific) by gavage for 7 days at 50 mg/kg/day ( n = 2 for each group). There was no binding to hepatic proteins from untreated control and AcHz-treated mice, whereas hepatic proteins from INH-treated mice showed a large number of bands modified with INH. (C) Antiserum was tested for specificity on Western blots by preincubation with INH at 200 μM or 2 mM for 1 h at 4 °C, which prevented binding to the INH-modified hepatic proteins. (D) Binding of the anti-INH serum to INH-modified liver proteins on Western blots was compared to that of the preimmune serum from the same animal.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Modification, Concentration Assay, Binding Assay, Mouse Assay, Western Blot

    24) Product Images from "Conjugated linoleic acid-mediated inflammation and insulin resistance in human adipocytes are attenuated by resveratrol *"

    Article Title: Conjugated linoleic acid-mediated inflammation and insulin resistance in human adipocytes are attenuated by resveratrol *

    Journal: Journal of Lipid Research

    doi: 10.1194/jlr.M800258-JLR200

    RSV blocks 10,12 CLA-mediated insulin resistance. Cultures of newly differentiated human adipocytes were serum starved for ∼24 h and then treated for either 48 h (A) or 24 h (B) with BSA vehicle or 50 μM 10,12 CLA in the absence (−)
    Figure Legend Snippet: RSV blocks 10,12 CLA-mediated insulin resistance. Cultures of newly differentiated human adipocytes were serum starved for ∼24 h and then treated for either 48 h (A) or 24 h (B) with BSA vehicle or 50 μM 10,12 CLA in the absence (−)

    Techniques Used:

    RSV attenuates delipidation by 10,12 CLA. Cultures of newly differentiated human adipocytes were treated in adipocyte media for 7 days with BSA vehicle or 50 μM 10,12 CLA in the absence (−) or presence (+) of 50 μM RSV (R). Fresh
    Figure Legend Snippet: RSV attenuates delipidation by 10,12 CLA. Cultures of newly differentiated human adipocytes were treated in adipocyte media for 7 days with BSA vehicle or 50 μM 10,12 CLA in the absence (−) or presence (+) of 50 μM RSV (R). Fresh

    Techniques Used:

    RSV inhibits 10,12 CLA suppression of peroxisome proliferator-activated receptor γ (PPARγ) activity. Cultures of newly differentiated human adipocytes were serum starved for ∼24 h and then treated for 24 h with BSA vehicle or 50
    Figure Legend Snippet: RSV inhibits 10,12 CLA suppression of peroxisome proliferator-activated receptor γ (PPARγ) activity. Cultures of newly differentiated human adipocytes were serum starved for ∼24 h and then treated for 24 h with BSA vehicle or 50

    Techniques Used: Activity Assay

    25) Product Images from "The Strategy of T Cell Antigen-presenting Cell Encounter in Antigen-draining Lymph Nodes Revealed by Imaging of Initial T Cell Activation"

    Article Title: The Strategy of T Cell Antigen-presenting Cell Encounter in Antigen-draining Lymph Nodes Revealed by Imaging of Initial T Cell Activation

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20030167

    CD4 + T cells cluster around Ag-loaded DCs in the outer paracortex of draining LNs. (a) Mice were primed by subcutaneous injection of BSA-coupled fluorescent microspheres. Draining LNs from five mice were harvested 24 h later and the expression of CD11c, CD11b, and CD8α was examined by FACS ® analysis of total LN cells or CD11c + cells, as indicated. (b–d) Mice were adoptively transferred with dye-labeled 3A9 CD4 T cells and then primed 24 h later with BSA- or HEL-coupled fluorescent microspheres, as indicated. Draining LNs were harvested 24 h (b and c) or 3 d (d) after immunization. (b) Clustering of 3A9 T cells (red) with microsphere-positive cells (green) near the B cell area revealed by anti-B220 staining (blue) is shown. (c) Phalloidin staining highlights the tight interaction between 3A9 T cells (diffuse green staining) and bead-containing DCs (red dots). (d) FACS ® analysis of CFSE profile of 3A9 T cells. One experiment out of two (a and c) or three (b and d) with similar results is presented.
    Figure Legend Snippet: CD4 + T cells cluster around Ag-loaded DCs in the outer paracortex of draining LNs. (a) Mice were primed by subcutaneous injection of BSA-coupled fluorescent microspheres. Draining LNs from five mice were harvested 24 h later and the expression of CD11c, CD11b, and CD8α was examined by FACS ® analysis of total LN cells or CD11c + cells, as indicated. (b–d) Mice were adoptively transferred with dye-labeled 3A9 CD4 T cells and then primed 24 h later with BSA- or HEL-coupled fluorescent microspheres, as indicated. Draining LNs were harvested 24 h (b and c) or 3 d (d) after immunization. (b) Clustering of 3A9 T cells (red) with microsphere-positive cells (green) near the B cell area revealed by anti-B220 staining (blue) is shown. (c) Phalloidin staining highlights the tight interaction between 3A9 T cells (diffuse green staining) and bead-containing DCs (red dots). (d) FACS ® analysis of CFSE profile of 3A9 T cells. One experiment out of two (a and c) or three (b and d) with similar results is presented.

    Techniques Used: Mouse Assay, Injection, Expressing, FACS, Labeling, Staining

    26) Product Images from "ADAP-SLP-76 Binding Differentially Regulates Supramolecular Activation Cluster (SMAC) Formation Relative to T Cell-APC Conjugation"

    Article Title: ADAP-SLP-76 Binding Differentially Regulates Supramolecular Activation Cluster (SMAC) Formation Relative to T Cell-APC Conjugation

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20040780

    ADAP–SLP-76 binding regulates integrin adhesion and clustering. GFP, ADAP-GFP, and M12-GFP–infected T8.1 cells were unstimulated or stimulated with 2C11 or PMA for 30 min and incubated on BSA, ICAM-1Fc–coated plates (a) or fibronectin (FN)-coated plates (b). (c) GFP, ADAP-GFP, and M12-GFP–infected T8.1 cells were stimulated with 2C11 for 30 min, fixed, and stained with anti-CD11a and Alexa Fluor 546–conjugated anti–rat antibody. Anti–LFA-1 capping was defined by the presence of a discrete polarized cap at one end of the cell. (bottom) Histogram showing the percentage of T8.1 cells with LFA-1 clustering. (d) GFP, ADAP-GFP, and M12-GFP–infected T8.1 cells were cocultured with Ttox pulsed L625.7 cells for 30 min, fixed, and stained for LFA-1 as before. Imaging and measurements of staining at the immunological synapse amongst individual conjugates ( > 45) were performed by laser-scanning microscopy. (top) DIC image (left) showed the conjugate and fluorescence image (right) showed LFA-1 clustering at the interface between T8.1 and L625.7 cells. (bottom) Histogram showed the percentage of LFA-1 at the interface relative to the total surface LFA-1.
    Figure Legend Snippet: ADAP–SLP-76 binding regulates integrin adhesion and clustering. GFP, ADAP-GFP, and M12-GFP–infected T8.1 cells were unstimulated or stimulated with 2C11 or PMA for 30 min and incubated on BSA, ICAM-1Fc–coated plates (a) or fibronectin (FN)-coated plates (b). (c) GFP, ADAP-GFP, and M12-GFP–infected T8.1 cells were stimulated with 2C11 for 30 min, fixed, and stained with anti-CD11a and Alexa Fluor 546–conjugated anti–rat antibody. Anti–LFA-1 capping was defined by the presence of a discrete polarized cap at one end of the cell. (bottom) Histogram showing the percentage of T8.1 cells with LFA-1 clustering. (d) GFP, ADAP-GFP, and M12-GFP–infected T8.1 cells were cocultured with Ttox pulsed L625.7 cells for 30 min, fixed, and stained for LFA-1 as before. Imaging and measurements of staining at the immunological synapse amongst individual conjugates ( > 45) were performed by laser-scanning microscopy. (top) DIC image (left) showed the conjugate and fluorescence image (right) showed LFA-1 clustering at the interface between T8.1 and L625.7 cells. (bottom) Histogram showed the percentage of LFA-1 at the interface relative to the total surface LFA-1.

    Techniques Used: Binding Assay, Infection, Incubation, Staining, Imaging, Laser-Scanning Microscopy, Fluorescence

    27) Product Images from "Apolipoprotein A-IV enhances cholecystokinin secretion"

    Article Title: Apolipoprotein A-IV enhances cholecystokinin secretion

    Journal: Physiology & behavior

    doi: 10.1016/j.physbeh.2018.01.019

    Comparison of CCK levels in the media of STC-1 cells after treated with Apo-AIV and BSA. STC-1 cells were incubated with vehicle, ApoA-IV proteins or BSA at 200 μg/ml, and the media were collected after a 2-h incubation for the determination of CCK level (pmol/mL equivalent to CCK-33). Data are expressed as mean ± SEM for the medium of STC-1 cells from 5-6 wells per group. Values with asterisks represent significant differences relative to the vehicle treatment (P
    Figure Legend Snippet: Comparison of CCK levels in the media of STC-1 cells after treated with Apo-AIV and BSA. STC-1 cells were incubated with vehicle, ApoA-IV proteins or BSA at 200 μg/ml, and the media were collected after a 2-h incubation for the determination of CCK level (pmol/mL equivalent to CCK-33). Data are expressed as mean ± SEM for the medium of STC-1 cells from 5-6 wells per group. Values with asterisks represent significant differences relative to the vehicle treatment (P

    Techniques Used: Incubation

    28) Product Images from "Inhibition of the Complement Membrane Attack Complex by Schistosoma mansoni Paramyosin "

    Article Title: Inhibition of the Complement Membrane Attack Complex by Schistosoma mansoni Paramyosin

    Journal: Infection and Immunity

    doi: 10.1128/IAI.71.11.6402-6410.2003

    Binding of paramyosin to C8 and C9. Purified human C8 and C9 and BSA (1 μg each) were subjected to SDS-8% PAGE and transferred onto a nitrocellulose membrane. After blocking with 5% milk in TBST, the membrane was incubated with recombinant paramyosin (4 μg/ml) in blocking solution for 2 h at 37°C. After two washes with TBST, the membrane was reacted with monoclonal anti-paramyosin antibody (1:4) (Anti-Pmy) or an isotype-matched, unrelated monoclonal antibody (Control) and then incubated with peroxidase-conjugated goat anti-mouse IgG (1:5,000) for 1 h at room temperature. Bands were visualized by enhanced chemiluminescence. Lane a, C8α and C8γ (upper band) and C8β (lower band); lane b, C9; lane c, BSA.
    Figure Legend Snippet: Binding of paramyosin to C8 and C9. Purified human C8 and C9 and BSA (1 μg each) were subjected to SDS-8% PAGE and transferred onto a nitrocellulose membrane. After blocking with 5% milk in TBST, the membrane was incubated with recombinant paramyosin (4 μg/ml) in blocking solution for 2 h at 37°C. After two washes with TBST, the membrane was reacted with monoclonal anti-paramyosin antibody (1:4) (Anti-Pmy) or an isotype-matched, unrelated monoclonal antibody (Control) and then incubated with peroxidase-conjugated goat anti-mouse IgG (1:5,000) for 1 h at room temperature. Bands were visualized by enhanced chemiluminescence. Lane a, C8α and C8γ (upper band) and C8β (lower band); lane b, C9; lane c, BSA.

    Techniques Used: Binding Assay, Purification, Polyacrylamide Gel Electrophoresis, Blocking Assay, Incubation, Recombinant

    Binding of paramyosin to blotted C9: competition with soluble C9 or CD59. (Lanes a and b) Purified human C9 (lanes a) or BSA (lanes b) (1 μg each) was blotted onto nitrocellulose membrane. The membrane was reacted with paramyosin (Pmy) or paramyosin preincubated with C9 (1:5 [wt/wt]) (Pmy+C9) or buffer (control) and then with monoclonal anti-paramyosin antibody, peroxidase-conjugated goat anti-mouse IgG, and enhanced chemiluminescence reagent. (Lanes c to e) Purified human C9 was blotted onto nitrocellulose membrane and reacted with recombinant paramyosin preincubated with or without purified human CD59. Further details are provided in Materials and Methods. Bound paramyosin was detected with monoclonal anti-paramyosin antibodies, goat anti-mouse IgG, and enhanced chemiluminescence reagent. Lane c, 2 μg of paramyosin without CD59; lane d, 2 μg of paramyosin plus 0.33 μg of CD59; lane e, 2 μg paramyosin plus 1 μg of CD59.
    Figure Legend Snippet: Binding of paramyosin to blotted C9: competition with soluble C9 or CD59. (Lanes a and b) Purified human C9 (lanes a) or BSA (lanes b) (1 μg each) was blotted onto nitrocellulose membrane. The membrane was reacted with paramyosin (Pmy) or paramyosin preincubated with C9 (1:5 [wt/wt]) (Pmy+C9) or buffer (control) and then with monoclonal anti-paramyosin antibody, peroxidase-conjugated goat anti-mouse IgG, and enhanced chemiluminescence reagent. (Lanes c to e) Purified human C9 was blotted onto nitrocellulose membrane and reacted with recombinant paramyosin preincubated with or without purified human CD59. Further details are provided in Materials and Methods. Bound paramyosin was detected with monoclonal anti-paramyosin antibodies, goat anti-mouse IgG, and enhanced chemiluminescence reagent. Lane c, 2 μg of paramyosin without CD59; lane d, 2 μg of paramyosin plus 0.33 μg of CD59; lane e, 2 μg paramyosin plus 1 μg of CD59.

    Techniques Used: Binding Assay, Purification, Recombinant

    29) Product Images from "Directly Coupled High-Performance Liquid Chromatography-Accelerator Mass Spectrometry Measurement of Chemically Modified Protein and Peptides"

    Article Title: Directly Coupled High-Performance Liquid Chromatography-Accelerator Mass Spectrometry Measurement of Chemically Modified Protein and Peptides

    Journal: Analytical chemistry

    doi: 10.1021/ac303609n

    Absolute quantitation of an albumin 14 C-iodoacetamide modification competitive binding experiment. Treated and untreated BSA samples were exposed to 1 pCi 14 C-iodoacetamide and described as follows: 14 C-iodoacetamide-modified native BSA ( 14 C-IAC-native):native
    Figure Legend Snippet: Absolute quantitation of an albumin 14 C-iodoacetamide modification competitive binding experiment. Treated and untreated BSA samples were exposed to 1 pCi 14 C-iodoacetamide and described as follows: 14 C-iodoacetamide-modified native BSA ( 14 C-IAC-native):native

    Techniques Used: Quantitation Assay, Modification, Binding Assay

    30) Product Images from "ToF-SIMS and XPS Characterization of Protein Films Adsorbed onto Bare and Sodium Styrene Sulfonate Grafted Gold Substrates"

    Article Title: ToF-SIMS and XPS Characterization of Protein Films Adsorbed onto Bare and Sodium Styrene Sulfonate Grafted Gold Substrates

    Journal: Langmuir : the ACS journal of surfaces and colloids

    doi: 10.1021/acs.langmuir.5b04743

    Amino acid peak list PCA results. Panels (A) and (B) are the scores and loadings plots for BSA. Panels (C) and (D) are the scores and loadings plots for Fgn. Panels (E) and (F) and the scores and loadings plots for IgG. In panels (A), (C), and (E),
    Figure Legend Snippet: Amino acid peak list PCA results. Panels (A) and (B) are the scores and loadings plots for BSA. Panels (C) and (D) are the scores and loadings plots for Fgn. Panels (E) and (F) and the scores and loadings plots for IgG. In panels (A), (C), and (E),

    Techniques Used:

    XPS isotherms for (A) BSA, (B) Fgn, (C) IgG, and (D) plasma adsorption onto gold and NaSS surfaces. Panels (E) through (H) are isotherms of the PC1 scores obtained from PCA using peak lists containing all peaks that are at least 3x the background intensity.
    Figure Legend Snippet: XPS isotherms for (A) BSA, (B) Fgn, (C) IgG, and (D) plasma adsorption onto gold and NaSS surfaces. Panels (E) through (H) are isotherms of the PC1 scores obtained from PCA using peak lists containing all peaks that are at least 3x the background intensity.

    Techniques Used: Adsorption

    ToF-SIMS peak intensity ratios reveal changes in adsorbed protein structure for (A) BSA and (B) IgG adsorbed onto gold and NaSS surfaces. Insets show the crystal structure of each protein. In (A) Asn and Tyr are highlighted in yellow while His is highlighted
    Figure Legend Snippet: ToF-SIMS peak intensity ratios reveal changes in adsorbed protein structure for (A) BSA and (B) IgG adsorbed onto gold and NaSS surfaces. Insets show the crystal structure of each protein. In (A) Asn and Tyr are highlighted in yellow while His is highlighted

    Techniques Used:

    ToF-SIMS peak intensity ratios suggest levels of denaturation of (A) BSA, (B) Fgn, (C) and IgG adsorbed onto gold and NaSS surfaces. Insets show the crystal structure of each protein. Bars represent mean amino acid peak ratios for proteins adsorbed from
    Figure Legend Snippet: ToF-SIMS peak intensity ratios suggest levels of denaturation of (A) BSA, (B) Fgn, (C) and IgG adsorbed onto gold and NaSS surfaces. Insets show the crystal structure of each protein. Bars represent mean amino acid peak ratios for proteins adsorbed from

    Techniques Used:

    PC models for gold (A and B) and NaSS (C and D) surfaces. The models were constructed using the amino acids peaks from 100 μg/ml plasma adsorption ToF-SIMS data. Single-component BSA ( ), Fgn ( ), and IgG ( ) spectra were projected into the models.
    Figure Legend Snippet: PC models for gold (A and B) and NaSS (C and D) surfaces. The models were constructed using the amino acids peaks from 100 μg/ml plasma adsorption ToF-SIMS data. Single-component BSA ( ), Fgn ( ), and IgG ( ) spectra were projected into the models.

    Techniques Used: Construct, Adsorption

    31) Product Images from "HIV type 1 Gag virus-like particle budding from spheroplasts of Saccharomycescerevisiae"

    Article Title: HIV type 1 Gag virus-like particle budding from spheroplasts of Saccharomycescerevisiae

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

    doi: 10.1073/pnas.082281199

    Time courses of VLP production. After removal of the cell wall, yeast spheroplasts expressing the full-length Gag protein were cultured at 30°C. At each time point, VLPs were purified from the culture medium by sucrose gradient centrifugation. ( A ) VLP fractions detected by Western blotting using anti-HIV-1 CA Ab. Lanes: M, prestained molecular weight markers; pre, before removal of the cell wall; 0, just after removal of the cell wall; and 0.5–16, hours of cultivation following the removal. ( B ) VLP yields estimated by quantitative Western blotting and Coomassie brilliant blue staining. The VLP yield at each time point was estimated by comparison to standard dilutions prepared with purified soluble Gag protein (for Western blotting) or BSA (for Coomassie brilliant blue staining).
    Figure Legend Snippet: Time courses of VLP production. After removal of the cell wall, yeast spheroplasts expressing the full-length Gag protein were cultured at 30°C. At each time point, VLPs were purified from the culture medium by sucrose gradient centrifugation. ( A ) VLP fractions detected by Western blotting using anti-HIV-1 CA Ab. Lanes: M, prestained molecular weight markers; pre, before removal of the cell wall; 0, just after removal of the cell wall; and 0.5–16, hours of cultivation following the removal. ( B ) VLP yields estimated by quantitative Western blotting and Coomassie brilliant blue staining. The VLP yield at each time point was estimated by comparison to standard dilutions prepared with purified soluble Gag protein (for Western blotting) or BSA (for Coomassie brilliant blue staining).

    Techniques Used: Expressing, Cell Culture, Purification, Gradient Centrifugation, Western Blot, Molecular Weight, Staining

    32) Product Images from "Fetuin-A/Albumin-Mineral Complexes Resembling Serum Calcium Granules and Putative Nanobacteria: Demonstration of a Dual Inhibition-Seeding Concept"

    Article Title: Fetuin-A/Albumin-Mineral Complexes Resembling Serum Calcium Granules and Putative Nanobacteria: Demonstration of a Dual Inhibition-Seeding Concept

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0008058

    Seeding of NB-like particles by boiled fetuin-A and albumin in metastable versus supersaturated medium. (A) Metastable medium: BSF and bovine serum albumin (BSA) were boiled for the time indicated on the left side. The boiled protein solutions were then diluted into DMEM, separately or together, to the final concentrations indicated on the top. A 650 was then monitored following inoculation (“Day 1”) and after incubation in cell culture conditions for 1 month, as indicated on the right. Inoculation of either untreated BSF or BSF that had been boiled for either 10, 30, or 120 min did not result in any turbidity changes after 1 month of incubation. In contrast, pre-boiled BSA produced either bell-shaped or linear turbidity changes depending on the amount of boiling time. When added together, pre-boiled BSF and BSA produced additive effects. (B) Supersaturated medium: BSF and BSA were added to medium as in (A) that was also inoculated with 1 mM each of CaCl 2 and NaH 2 PO 4 . Notice the various patterns of turbidity changes associated with the three protein combinations. See the text for further details.
    Figure Legend Snippet: Seeding of NB-like particles by boiled fetuin-A and albumin in metastable versus supersaturated medium. (A) Metastable medium: BSF and bovine serum albumin (BSA) were boiled for the time indicated on the left side. The boiled protein solutions were then diluted into DMEM, separately or together, to the final concentrations indicated on the top. A 650 was then monitored following inoculation (“Day 1”) and after incubation in cell culture conditions for 1 month, as indicated on the right. Inoculation of either untreated BSF or BSF that had been boiled for either 10, 30, or 120 min did not result in any turbidity changes after 1 month of incubation. In contrast, pre-boiled BSA produced either bell-shaped or linear turbidity changes depending on the amount of boiling time. When added together, pre-boiled BSF and BSA produced additive effects. (B) Supersaturated medium: BSF and BSA were added to medium as in (A) that was also inoculated with 1 mM each of CaCl 2 and NaH 2 PO 4 . Notice the various patterns of turbidity changes associated with the three protein combinations. See the text for further details.

    Techniques Used: Incubation, Cell Culture, Produced

    33) Product Images from "A General Strategy for Generating Gradients of Bioactive Proteins on Electrospun Nanofiber Mats by Masking with Bovine Serum Albumin"

    Article Title: A General Strategy for Generating Gradients of Bioactive Proteins on Electrospun Nanofiber Mats by Masking with Bovine Serum Albumin

    Journal: Journal of materials chemistry. B, Materials for biology and medicine

    doi: 10.1039/C7TB00974G

    SEM images of random PCL nanofibers (A) before and (B–;D) after plasma treatment for 2 min, followed by soaking in (B) PBS, (C) 0.1% BSA solution, and (D) 0.5% BSA solution, respectively, for 1 h. The morphology and texture of the nanofibers are essentially preserved during the treatments.
    Figure Legend Snippet: SEM images of random PCL nanofibers (A) before and (B–;D) after plasma treatment for 2 min, followed by soaking in (B) PBS, (C) 0.1% BSA solution, and (D) 0.5% BSA solution, respectively, for 1 h. The morphology and texture of the nanofibers are essentially preserved during the treatments.

    Techniques Used:

    (A) Plots of the relative fluorescence intensities for the BSA-FITC adsorbed on the nanofibers as a function of soaking time in the 0.1% and 0.5% BSA solutions, respectively. The corresponding representative fluorescence micrographs of the nanofibers are also included (upper row: 0.1% BSA, and lower row: 0.5%). The extent of BSA adsorbed on PCL nanofibers is dependent on the duration of exposure to BSA solution (n=3 for each time point). The BSA concentration also affects the rate at which BSA adsorbs onto the nanofibers. (B) Representative fluorescence micrographs and their corresponding relative fluorescence intensities at different positions of nanofiber strips by varying the duration of contact time with a 0.1% BSA solution over a distance of 35 mm (n=9 at each position). A gradient in BSA can be clearly seen across the strip of nanofibers.
    Figure Legend Snippet: (A) Plots of the relative fluorescence intensities for the BSA-FITC adsorbed on the nanofibers as a function of soaking time in the 0.1% and 0.5% BSA solutions, respectively. The corresponding representative fluorescence micrographs of the nanofibers are also included (upper row: 0.1% BSA, and lower row: 0.5%). The extent of BSA adsorbed on PCL nanofibers is dependent on the duration of exposure to BSA solution (n=3 for each time point). The BSA concentration also affects the rate at which BSA adsorbs onto the nanofibers. (B) Representative fluorescence micrographs and their corresponding relative fluorescence intensities at different positions of nanofiber strips by varying the duration of contact time with a 0.1% BSA solution over a distance of 35 mm (n=9 at each position). A gradient in BSA can be clearly seen across the strip of nanofibers.

    Techniques Used: Fluorescence, Concentration Assay, Stripping Membranes

    34) Product Images from "Mass spectrometry-based proteomics of oxidative stress: Identification of 4-hydroxy-2-nonenal (HNE) adducts of amino acids using lysozyme and bovine serum albumin as model proteins"

    Article Title: Mass spectrometry-based proteomics of oxidative stress: Identification of 4-hydroxy-2-nonenal (HNE) adducts of amino acids using lysozyme and bovine serum albumin as model proteins

    Journal: Electrophoresis

    doi: 10.1002/elps.201600134

    Example of pyrrole-type His modification by HNE in BSA, at the peptide and amino acid levels. (A) XIC of peak with m/z 713.26 (2+) in BSA sample treated with 0, 2, 5 and 10 mM HNE. (B) Summed spectra from the XIC of the peak with m/z 713.26 (2+). (C) Product ions (MS/MS fragmentation) of the precursor ion from HNE-treated BSA, with m/z 713.26 (2+).
    Figure Legend Snippet: Example of pyrrole-type His modification by HNE in BSA, at the peptide and amino acid levels. (A) XIC of peak with m/z 713.26 (2+) in BSA sample treated with 0, 2, 5 and 10 mM HNE. (B) Summed spectra from the XIC of the peak with m/z 713.26 (2+). (C) Product ions (MS/MS fragmentation) of the precursor ion from HNE-treated BSA, with m/z 713.26 (2+).

    Techniques Used: Modification, Mass Spectrometry

    Example of HNE modification of Arg residue in BSA, at the peptide and amino acid levels. (A) Extracted ion chromatograms (XIC) of the peak with m/z 651.31 (2+) in BSA sample treated with 0, 2, 5 and 10 mM HNE. (B) Summed spectra from the XIC of the peak with m/z 651.31 (2+). (C) Product ions (MS/MS fragmentation) of the precursor ion from HNE-treated BSA, with m/z 651.31 (2+).
    Figure Legend Snippet: Example of HNE modification of Arg residue in BSA, at the peptide and amino acid levels. (A) Extracted ion chromatograms (XIC) of the peak with m/z 651.31 (2+) in BSA sample treated with 0, 2, 5 and 10 mM HNE. (B) Summed spectra from the XIC of the peak with m/z 651.31 (2+). (C) Product ions (MS/MS fragmentation) of the precursor ion from HNE-treated BSA, with m/z 651.31 (2+).

    Techniques Used: Modification, Mass Spectrometry

    Example of HNE modification of Thr residue in BSA, at the peptide and amino acid levels. (A) XIC of the peak with m/z 990.45 (2+) in BSA sample treated with 0, 2, 5 and 10 mM HNE. (B) Summed spectra from the XIC of the peak with m/z 990.45 (2+). (C) Product ions (MS/MS fragmentation) of the precursor ion from HNE-treated BSA, with m/z 990.45 (2+).
    Figure Legend Snippet: Example of HNE modification of Thr residue in BSA, at the peptide and amino acid levels. (A) XIC of the peak with m/z 990.45 (2+) in BSA sample treated with 0, 2, 5 and 10 mM HNE. (B) Summed spectra from the XIC of the peak with m/z 990.45 (2+). (C) Product ions (MS/MS fragmentation) of the precursor ion from HNE-treated BSA, with m/z 990.45 (2+).

    Techniques Used: Modification, Mass Spectrometry

    35) Product Images from "Increasing Dietary Medium-Chain Fatty Acid Ratio Mitigates High-fat Diet-Induced Non-Alcoholic Steatohepatitis by Regulating Autophagy"

    Article Title: Increasing Dietary Medium-Chain Fatty Acid Ratio Mitigates High-fat Diet-Induced Non-Alcoholic Steatohepatitis by Regulating Autophagy

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-14376-y

    Increasing MCFA ratio attenuated LCFAs-induced fat accumulation, insulin resistance, autophagy impairment, ER stress, and apoptosis in HepG2 cells. To investigate whether MCFAs can directly exert their protective effects in hepatic cells, the fat accumulation, insulin resistance, autophagy impairment, ER stress, and apoptosis were evaluated in fat-loaded HepG2 cells. For intracellular lipid quantification, 48-hour fat-loaded cells were fixed then stained with Nile Red and Hoechst 33342 ( A ). For the insulin sensitivity test, cells were incubated with 100 nM insulin for an additional 10 min after a 48-hour fatty acid treatment. Representative immunoblots and densitometric quantification of phospho-Akt (Ser473) ( B ), and the calculated insulin-stimulated Akt phosphorylation fold change results ( C ) are presented. Representative SQSTM1/p62 and LC3 immunoblots and densitometric results show that increasing MCFA/LCFA ratio protected HepG2 cells against LCFA-induced autophagy impairment ( D ). Autophagic flux changes were also confirmed by analyzing Bafilomycin A1 (50 nM, 3 hours)-induced LC3 II protein accumulation in BSA or fatty acid-treated HepG2 cells ( E ). Western blotting results of CHOP and cleaved caspase 3 show that MCFAs also attenuated LCFAs-induced ER stress and apoptosis in HepG2 cells ( F ). For densitometric analyses of Western blotting data, β-actin or total Akt (for Akt phosphorylation analysis) were used as the loading control. Values are mean ± SEM (n = 3). *, # (vs. BSA treated cells without insulin stimulation), and † (vs. SDF-treated cells without insulin stimulation) indicate statistical significance, P
    Figure Legend Snippet: Increasing MCFA ratio attenuated LCFAs-induced fat accumulation, insulin resistance, autophagy impairment, ER stress, and apoptosis in HepG2 cells. To investigate whether MCFAs can directly exert their protective effects in hepatic cells, the fat accumulation, insulin resistance, autophagy impairment, ER stress, and apoptosis were evaluated in fat-loaded HepG2 cells. For intracellular lipid quantification, 48-hour fat-loaded cells were fixed then stained with Nile Red and Hoechst 33342 ( A ). For the insulin sensitivity test, cells were incubated with 100 nM insulin for an additional 10 min after a 48-hour fatty acid treatment. Representative immunoblots and densitometric quantification of phospho-Akt (Ser473) ( B ), and the calculated insulin-stimulated Akt phosphorylation fold change results ( C ) are presented. Representative SQSTM1/p62 and LC3 immunoblots and densitometric results show that increasing MCFA/LCFA ratio protected HepG2 cells against LCFA-induced autophagy impairment ( D ). Autophagic flux changes were also confirmed by analyzing Bafilomycin A1 (50 nM, 3 hours)-induced LC3 II protein accumulation in BSA or fatty acid-treated HepG2 cells ( E ). Western blotting results of CHOP and cleaved caspase 3 show that MCFAs also attenuated LCFAs-induced ER stress and apoptosis in HepG2 cells ( F ). For densitometric analyses of Western blotting data, β-actin or total Akt (for Akt phosphorylation analysis) were used as the loading control. Values are mean ± SEM (n = 3). *, # (vs. BSA treated cells without insulin stimulation), and † (vs. SDF-treated cells without insulin stimulation) indicate statistical significance, P

    Techniques Used: Staining, Incubation, Western Blot

    36) Product Images from "Subnuclear Trafficking of Glucocorticoid Receptors In Vitro: Chromatin Recycling and Nuclear Export"

    Article Title: Subnuclear Trafficking of Glucocorticoid Receptors In Vitro: Chromatin Recycling and Nuclear Export

    Journal: The Journal of Cell Biology

    doi:

    In vitro nuclear export of GR is an ATP-driven process. Hormone-withdrawn GrH2 cells were permeabilized and then incubated with 10 mg/ml of BSA in transport buffer containing 20 mM sodium molybdate at 30°C with the following additions: ( A ) 4 mM ATP; ( B ) 4 mM GTP; ( C ) 4 mM each of ATP and GTPγS; ( D ) 4 mM each of GTP and AMP-PNP. GR in fixed cells was detected by IIF.
    Figure Legend Snippet: In vitro nuclear export of GR is an ATP-driven process. Hormone-withdrawn GrH2 cells were permeabilized and then incubated with 10 mg/ml of BSA in transport buffer containing 20 mM sodium molybdate at 30°C with the following additions: ( A ) 4 mM ATP; ( B ) 4 mM GTP; ( C ) 4 mM each of ATP and GTPγS; ( D ) 4 mM each of GTP and AMP-PNP. GR in fixed cells was detected by IIF.

    Techniques Used: In Vitro, Incubation

    In vitro nuclear export of GR stimulated by tyrosine phosphatase inhibitors is blocked by the tyrosine kinase inhibitors. Permeabilized, hormone-withdrawn GrH2 cells were incubated with BSA in transport buffer containing 4 mM ATP for 20 min at 30°C, in the absence ( A , C , E , G , and H ) or presence ( B , D , and F ) of 0.2 mM genistein. 20 mM sodium molybdate was included in A , B , and H , 7.5 mM sodium vanadate in C and D , 50 μg/ml of heparin in E and F , 1 μM microcystin in G , and 1 mM tyrphostin AG126 ( AG 126 ) in H. GR was detected in fixed cells by IIF.
    Figure Legend Snippet: In vitro nuclear export of GR stimulated by tyrosine phosphatase inhibitors is blocked by the tyrosine kinase inhibitors. Permeabilized, hormone-withdrawn GrH2 cells were incubated with BSA in transport buffer containing 4 mM ATP for 20 min at 30°C, in the absence ( A , C , E , G , and H ) or presence ( B , D , and F ) of 0.2 mM genistein. 20 mM sodium molybdate was included in A , B , and H , 7.5 mM sodium vanadate in C and D , 50 μg/ml of heparin in E and F , 1 μM microcystin in G , and 1 mM tyrphostin AG126 ( AG 126 ) in H. GR was detected in fixed cells by IIF.

    Techniques Used: In Vitro, Incubation

    ( a ) Western blot analysis of molybdate-stimulated in vitro GR nuclear export. (Lanes 1–8 ) GrH2 cells treated with corticosterone ( Cort ) for 1 h and then withdrawn from hormone for 20 min. (Lanes 9–12 ) GrH2 cells treated with corticosterone for 80 min. Harvested cells were permeabilized in suspension, and aliquots of intact nuclei were incubated with 50 μl of reaction mixture containing 10 mg/ml BSA in transport buffer, 4 mM ATP (lanes 1 , 2 , 5 , 6 , 9 , and 10 ), or 4 mM GTP (lanes 3 , 7 , and 11 ) with an energy-regenerating system, and 20 mM sodium molybdate where indicated (lanes 2 , 3 , 4 , 6 , 7 , 8 , 10 , 11 , and 12 ). After a 20min incubation at 30°C, each nuclear suspension was split into two identical samples. One sample was subjected to SDS-PAGE directly (lanes 1–4 , Whole ). The other identical sample was washed, and nuclei were recovered and subjected to SDS-PAGE for detection of the remaining nuclear GR (lanes 5–12 , Nuc Prep ). ( b ) Quantification of GR levels observed in Western blots by densitometry. GR levels were normalized to the internal control NuMA protein. (Bars 1–8 ) Average of four experiments; (bars 9–12 ) average of two experiments. Whole , whole reaction mix; Nuc Prep , nuclear pellets.
    Figure Legend Snippet: ( a ) Western blot analysis of molybdate-stimulated in vitro GR nuclear export. (Lanes 1–8 ) GrH2 cells treated with corticosterone ( Cort ) for 1 h and then withdrawn from hormone for 20 min. (Lanes 9–12 ) GrH2 cells treated with corticosterone for 80 min. Harvested cells were permeabilized in suspension, and aliquots of intact nuclei were incubated with 50 μl of reaction mixture containing 10 mg/ml BSA in transport buffer, 4 mM ATP (lanes 1 , 2 , 5 , 6 , 9 , and 10 ), or 4 mM GTP (lanes 3 , 7 , and 11 ) with an energy-regenerating system, and 20 mM sodium molybdate where indicated (lanes 2 , 3 , 4 , 6 , 7 , 8 , 10 , 11 , and 12 ). After a 20min incubation at 30°C, each nuclear suspension was split into two identical samples. One sample was subjected to SDS-PAGE directly (lanes 1–4 , Whole ). The other identical sample was washed, and nuclei were recovered and subjected to SDS-PAGE for detection of the remaining nuclear GR (lanes 5–12 , Nuc Prep ). ( b ) Quantification of GR levels observed in Western blots by densitometry. GR levels were normalized to the internal control NuMA protein. (Bars 1–8 ) Average of four experiments; (bars 9–12 ) average of two experiments. Whole , whole reaction mix; Nuc Prep , nuclear pellets.

    Techniques Used: Western Blot, In Vitro, Incubation, SDS Page

    Molybdate-stimulated in vitro nuclear export of VAN556 GR and hsp90. CHO cells stably transfected with VAN556 GR were permeabilized and then incubated with 50 μl of 10 mg/ml BSA in transport buffer for 20 min at 30°C with the following additions: ( A and B ) 4 mM ATP; ( C and D ) 4 mM ATP and 20 mM sodium molybdate; ( E and F ) 20 mM sodium molybdate. GR ( A , C , and E ) and hsp90 ( B , D , and F ) were detected in fixed cells by costaining with the BuGR2 mAb and TSTA rabbit serum.
    Figure Legend Snippet: Molybdate-stimulated in vitro nuclear export of VAN556 GR and hsp90. CHO cells stably transfected with VAN556 GR were permeabilized and then incubated with 50 μl of 10 mg/ml BSA in transport buffer for 20 min at 30°C with the following additions: ( A and B ) 4 mM ATP; ( C and D ) 4 mM ATP and 20 mM sodium molybdate; ( E and F ) 20 mM sodium molybdate. GR ( A , C , and E ) and hsp90 ( B , D , and F ) were detected in fixed cells by costaining with the BuGR2 mAb and TSTA rabbit serum.

    Techniques Used: In Vitro, Stable Transfection, Transfection, Incubation

    Stimulation of in vitro GR nuclear export by sodium molybdate. Hormone-withdrawn GrH2 cells ( A–E , G , and H ) were permeabilized and then incubated with 10 mg/ml of BSA in transport buffer at 30°C ( A , B , and D–H ) or 0°C ( C ) with the following additions: ( A ) ATP; ( B and C ) ATP and 20 mM sodium molybdate; ( D ) 20 mM sodium molybdate; ( E ) 0.2 mg/ml WGA, ATP, and 20 mM sodium molybdate; ( F ) corticosterone-treated GrH2 cells were permeabilized and then incubated for 20 min at 30°C with ATP and 20 mM sodium molybdate; ( G ) ATP, 20 mM sodium molybdate, and the Ab-1 anti-NuMA antibody; and ( H ) ATP and 20 mM sodium molybdate. In A–F , cells were fixed with methanol and subjected to IIF to detect GR. In G , permeabilized cells were fixed after the incubation described above and treated with a FITC-coupled secondary antibody to detect the Ab-1 anti-NuMA primary antibody. In H , ATP- and molybdate-treated permeabilized cells were fixed and then subjected to IIF with the Ab-1 antibody to detect nuclear NuMA.
    Figure Legend Snippet: Stimulation of in vitro GR nuclear export by sodium molybdate. Hormone-withdrawn GrH2 cells ( A–E , G , and H ) were permeabilized and then incubated with 10 mg/ml of BSA in transport buffer at 30°C ( A , B , and D–H ) or 0°C ( C ) with the following additions: ( A ) ATP; ( B and C ) ATP and 20 mM sodium molybdate; ( D ) 20 mM sodium molybdate; ( E ) 0.2 mg/ml WGA, ATP, and 20 mM sodium molybdate; ( F ) corticosterone-treated GrH2 cells were permeabilized and then incubated for 20 min at 30°C with ATP and 20 mM sodium molybdate; ( G ) ATP, 20 mM sodium molybdate, and the Ab-1 anti-NuMA antibody; and ( H ) ATP and 20 mM sodium molybdate. In A–F , cells were fixed with methanol and subjected to IIF to detect GR. In G , permeabilized cells were fixed after the incubation described above and treated with a FITC-coupled secondary antibody to detect the Ab-1 anti-NuMA primary antibody. In H , ATP- and molybdate-treated permeabilized cells were fixed and then subjected to IIF with the Ab-1 antibody to detect nuclear NuMA.

    Techniques Used: In Vitro, Incubation, Whole Genome Amplification

    Selective effects of tungstate on in vitro nuclear protein export. Permeabilized HeLa cells were incubated for 20 min at 30°C with BSA in transport buffer containing 20 mM sodium tungstate, in the absence ( A , C , E , and G ) or presence of 4 mM ATP ( B , D , F , and H ). Fixed cells were subjected to costaining to detect either hnRNP A1 ( A and B ) and hsp56 ( C and D ), or hnRNP C ( G and H ) and hsp90 ( E and F ) in the same fields of cells.
    Figure Legend Snippet: Selective effects of tungstate on in vitro nuclear protein export. Permeabilized HeLa cells were incubated for 20 min at 30°C with BSA in transport buffer containing 20 mM sodium tungstate, in the absence ( A , C , E , and G ) or presence of 4 mM ATP ( B , D , F , and H ). Fixed cells were subjected to costaining to detect either hnRNP A1 ( A and B ) and hsp56 ( C and D ), or hnRNP C ( G and H ) and hsp90 ( E and F ) in the same fields of cells.

    Techniques Used: In Vitro, Incubation

    37) Product Images from "Identification of EBP50: A PDZ-containing Phosphoprotein that Associates with Members of the Ezrin-Radixin-Moesin Family "

    Article Title: Identification of EBP50: A PDZ-containing Phosphoprotein that Associates with Members of the Ezrin-Radixin-Moesin Family

    Journal: The Journal of Cell Biology

    doi:

    Identification of N-ERMAD binding proteins. ( A ) Lysis buffer, or placental extracts, or brain extracts were mixed with BSA–agarose ( BSA ), ezrin–N-ERMAD-agarose ( E-N ), or moesin– N-ERMAD-agarose ( M-N ), and then were washed extensively in 0.5 M NaCl and bound proteins eluted and resolved on a 6–20% silver stained gradient SDS gel. For some reactions, a fourfold molar excess of competitor ezrin N-ERMAD (+ C ) or mock competitor BSA (+ mock ) was added to the extract before mixing with moesin–N-ERMAD-agarose. ( B ) One quarter the amount of the same samples shown in A were resolved on a 10% gel, transferred to PVDF, and probed with biotinylated ezrin– N-ERMAD. Brackets indicate the N-ERMAD binding proteins, and arrowheads indicate ezrin. The mobilities of standard proteins are indicated in kD. DF , dye front.
    Figure Legend Snippet: Identification of N-ERMAD binding proteins. ( A ) Lysis buffer, or placental extracts, or brain extracts were mixed with BSA–agarose ( BSA ), ezrin–N-ERMAD-agarose ( E-N ), or moesin– N-ERMAD-agarose ( M-N ), and then were washed extensively in 0.5 M NaCl and bound proteins eluted and resolved on a 6–20% silver stained gradient SDS gel. For some reactions, a fourfold molar excess of competitor ezrin N-ERMAD (+ C ) or mock competitor BSA (+ mock ) was added to the extract before mixing with moesin–N-ERMAD-agarose. ( B ) One quarter the amount of the same samples shown in A were resolved on a 10% gel, transferred to PVDF, and probed with biotinylated ezrin– N-ERMAD. Brackets indicate the N-ERMAD binding proteins, and arrowheads indicate ezrin. The mobilities of standard proteins are indicated in kD. DF , dye front.

    Techniques Used: Binding Assay, Lysis, Staining, SDS-Gel

    38) Product Images from "Highly sensitive color-indicating and quantitative biosensor based on cholesteric liquid crystal"

    Article Title: Highly sensitive color-indicating and quantitative biosensor based on cholesteric liquid crystal

    Journal: Biomedical Optics Express

    doi: 10.1364/BOE.6.005033

    POM images of VAC cells prepared with various concentrations (0 to 1 mg/ml) of BSA immobilized atop the DMOAP layers.
    Figure Legend Snippet: POM images of VAC cells prepared with various concentrations (0 to 1 mg/ml) of BSA immobilized atop the DMOAP layers.

    Techniques Used:

    39) Product Images from "Advanced glycation end products induce the expression of interleukin-6 and interleukin-8 by receptor for advanced glycation end product-mediated activation of mitogen-activated protein kinases and nuclear factor-?B in human osteoarthritis chondrocytes"

    Article Title: Advanced glycation end products induce the expression of interleukin-6 and interleukin-8 by receptor for advanced glycation end product-mediated activation of mitogen-activated protein kinases and nuclear factor-?B in human osteoarthritis chondrocytes

    Journal: Rheumatology (Oxford, England)

    doi: 10.1093/rheumatology/keq380

    Activation of RAGE-mediated NF-κB signalling by AGE-BSA in primary human OA chondrocytes. ( a ) Primary chondrocytes were pre-treated with sRAGE (200 µg/ml) for 2 h prior to AGE-BSA (50 µg/ml) stimulation or ( b ) RAGE-siRNA-transfected
    Figure Legend Snippet: Activation of RAGE-mediated NF-κB signalling by AGE-BSA in primary human OA chondrocytes. ( a ) Primary chondrocytes were pre-treated with sRAGE (200 µg/ml) for 2 h prior to AGE-BSA (50 µg/ml) stimulation or ( b ) RAGE-siRNA-transfected

    Techniques Used: Activation Assay, Transfection

    Activation of NF-κB is required for AGE-BSA-induced expression of IL-6 and IL-8 in primary human OA chondrocytes. Primary human OA chondrocytes were pre-treated with inhibitors specific for IKKα/β (Bay 11-7082), IKKβ (parthenolide),
    Figure Legend Snippet: Activation of NF-κB is required for AGE-BSA-induced expression of IL-6 and IL-8 in primary human OA chondrocytes. Primary human OA chondrocytes were pre-treated with inhibitors specific for IKKα/β (Bay 11-7082), IKKβ (parthenolide),

    Techniques Used: Activation Assay, Expressing

    Different MAPKs regulate AGE-BSA-induced expression of IL-6 and IL-8 in primary human OA chondrocytes. Effect of MAPK inhibition on the gene expression of IL-6 ( a ) and IL-8 ( b ) in AGE-BSA-stimulated OA chondrocytes. Primary human OA chondrocytes were
    Figure Legend Snippet: Different MAPKs regulate AGE-BSA-induced expression of IL-6 and IL-8 in primary human OA chondrocytes. Effect of MAPK inhibition on the gene expression of IL-6 ( a ) and IL-8 ( b ) in AGE-BSA-stimulated OA chondrocytes. Primary human OA chondrocytes were

    Techniques Used: Expressing, Inhibition

    Activation of RAGE-mediated MAPK signalling by AGE-BSA in primary human OA chondrocytes. Effect of sRAGE ( a ) and RAGE knockdown ( b ) on MAPK phosphorylation in AGE-BSA-stimulated OA chondrocytes. After pre-treatment with sRAGE (200 µg/ml)
    Figure Legend Snippet: Activation of RAGE-mediated MAPK signalling by AGE-BSA in primary human OA chondrocytes. Effect of sRAGE ( a ) and RAGE knockdown ( b ) on MAPK phosphorylation in AGE-BSA-stimulated OA chondrocytes. After pre-treatment with sRAGE (200 µg/ml)

    Techniques Used: Activation Assay

    AGE-BSA or S100A4 enhances the production of IL-6 and IL-8 in human OA cartilage explants. Human OA cartilage explants in serum-free DMEM were stimulated with AGE-BSA or S100A4 protein for 24 h at 37°C. Production of IL-6 ( a, b ) and IL-8
    Figure Legend Snippet: AGE-BSA or S100A4 enhances the production of IL-6 and IL-8 in human OA cartilage explants. Human OA cartilage explants in serum-free DMEM were stimulated with AGE-BSA or S100A4 protein for 24 h at 37°C. Production of IL-6 ( a, b ) and IL-8

    Techniques Used:

    Enhanced production of IL-6 and IL-8 by AGE-BSA-stimulated primary human OA chondrocytes. Effect of AGE-BSA on the protein production of IL-6 ( a ) and IL-8 ( b ) in primary human OA chondrocytes. Primary human OA chondrocytes were treated with AGE-BSA (5–100 µg/ml)
    Figure Legend Snippet: Enhanced production of IL-6 and IL-8 by AGE-BSA-stimulated primary human OA chondrocytes. Effect of AGE-BSA on the protein production of IL-6 ( a ) and IL-8 ( b ) in primary human OA chondrocytes. Primary human OA chondrocytes were treated with AGE-BSA (5–100 µg/ml)

    Techniques Used:

    Expression of IL-6 and IL-8 in AGE-BSA-stimulated primary human OA chondrocytes. Effect of AGE-BSA on the gene expression of IL-6 ( a ) and IL-8 ( b ) in primary human OA chondrocytes. Primary human OA chondrocytes were treated with AGE-BSA (5–100 µg/ml)
    Figure Legend Snippet: Expression of IL-6 and IL-8 in AGE-BSA-stimulated primary human OA chondrocytes. Effect of AGE-BSA on the gene expression of IL-6 ( a ) and IL-8 ( b ) in primary human OA chondrocytes. Primary human OA chondrocytes were treated with AGE-BSA (5–100 µg/ml)

    Techniques Used: Expressing

    Effect of sRAGE or RAGE-knockdown on the AGE-BSA or S100A4 protein-induced expression of IL-6 and IL-8 in primary human OA chondrocytes. Primary OA chondrocytes were pre-treated with sRAGE (200 µg/ml) for 2 h and then stimulated
    Figure Legend Snippet: Effect of sRAGE or RAGE-knockdown on the AGE-BSA or S100A4 protein-induced expression of IL-6 and IL-8 in primary human OA chondrocytes. Primary OA chondrocytes were pre-treated with sRAGE (200 µg/ml) for 2 h and then stimulated

    Techniques Used: Expressing

    40) Product Images from "Lyn Facilitates Glioblastoma Cell Survival under Conditions of Nutrient Deprivation by Promoting Autophagy"

    Article Title: Lyn Facilitates Glioblastoma Cell Survival under Conditions of Nutrient Deprivation by Promoting Autophagy

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0070804

    Expression of CA-Lyn promotes the survival of nutrient-deprived U87 human GBM cells. U87 human GBM cells expressing CA-Lyn, DN-Lyn or LV control were plated in DMEM with L-glutamine and 10% FBS. After 12 h, nutrient deprivation was induced by replacing the medium with L-glutamine- and FBS-free DMEM with 1% BSA. A, After 48 h of nutrient deprivation, whole cell lysates were western blotted with the indicated antibodies. B-D, After 7 days of nutrient deprivation, cell viability was determined by trypan blue exclusion (B–D) and apoptosis by FACs analysis of Annexin-V-labeled cells (E), blotting for cleaved caspase-7 (F), or FACS analysis for Annexin-V and propidium-iodide-labeled cells (G). B–E, and G, Conditions were assayed in replicas of 3 or 4, and the data analyzed and plotted as the mean±SEM. D E, Statistical analysis using one-way ANOVA. G, Statistical analysis using t-test.
    Figure Legend Snippet: Expression of CA-Lyn promotes the survival of nutrient-deprived U87 human GBM cells. U87 human GBM cells expressing CA-Lyn, DN-Lyn or LV control were plated in DMEM with L-glutamine and 10% FBS. After 12 h, nutrient deprivation was induced by replacing the medium with L-glutamine- and FBS-free DMEM with 1% BSA. A, After 48 h of nutrient deprivation, whole cell lysates were western blotted with the indicated antibodies. B-D, After 7 days of nutrient deprivation, cell viability was determined by trypan blue exclusion (B–D) and apoptosis by FACs analysis of Annexin-V-labeled cells (E), blotting for cleaved caspase-7 (F), or FACS analysis for Annexin-V and propidium-iodide-labeled cells (G). B–E, and G, Conditions were assayed in replicas of 3 or 4, and the data analyzed and plotted as the mean±SEM. D E, Statistical analysis using one-way ANOVA. G, Statistical analysis using t-test.

    Techniques Used: Expressing, Western Blot, FACS, Labeling

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    Article Snippet: .. The day after, PHHs were overnight inoculated with 2,500 genome equivalents (GE) per cell of cell culture-derived HBV (genotype D) in PHH cultivation medium (Dulbecco's modified Eagle medium [DMEM]) containing 10% fetal bovine serum (FBS) and 2% dimethyl sulfoxide (DMSO) supplemented with HEPES, l -proline, insulin, epidermal growth factor, dexamethasone, and ascorbic acid-2-phosphate (purchased from Life Technologies and Sigma) without additional plasma proteins (this medium is referred to as HCM without plasma proteins) or in hepatocyte cultivation medium (HCM) spiked with either 4.3% (43 g/liter) bovine serum albumin (BSA) (Sigma) and 0.07% (0.7 g/liter) human alpha-1 acid glycoprotein (hA1GP) (Sigma) or with 8% BSA (80 g/liter) and 0.07% hA1GP. .. For inoculation, PHH medium was supplemented with 4% polyethylene glycol (PEG) 8000 (Sigma).

    Incubation:

    Article Title: Ligand-modified gene carriers increased uptake in target cells but reduced DNA release and transfection efficiency
    Article Snippet: .. Coumarin-6 loaded particles or BSA-FITC (Sigma) was delivered through incubation for 6 hrs prior to fixation. .. Next, coverslips were washed twice with 1× PBS and fixed with 4% Paraformaldehyde (Fisher Scientific).

    SDS Page:

    Article Title: Detection of ubiquityl-calmodulin conjugates with a novel high-molecular weight ubiquitylprotein-isopeptidase in rabbit tissues
    Article Snippet: .. The molecular weight standards for SDS-PAGE bovine serum albumin (BSA) 66 kDa, ovalbumin 45 kDa, glyceraldehyde 3-phosphate dehydrogenase 36 kDa, carbonic anhydratase 29.2 kDa, trypsinogen 25 kDa, trypsin inhibitor 20.1 kDa and lactalbumin 14.2 kDa were obtained from Sigma (Munich). ..

    Molecular Weight:

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  • 99
    Millipore anti gst antibody
    CPP-MT and CPP-MT-AuNP translocate intact human sperm. Notes: Sperm were incubated with 20 µg/mL of CPP-MT ( A ), CPP-MT-AuNP ( B ), or non-permeable <t>GST-Rab3A</t> ( C , control) for 3 hours at 37°C in HTF medium. Afterward, cells were treated with trypsin to eliminate external probe. Then, cells were fixed and spotted on glass slides and labeled with <t>Cy3-coupled</t> anti-GST antibody. At least 100 cells were quantified, and the population of sperm showing positive label was recorded. The bars represent mean values ± SEM, calculated from three independent experiments. The images are representative of the experimental conditions described. One-way ANOVA shows statistically significant difference between relative labeled cells percentages incubated with CPP-MT ( A ′) or CPP-MT-AuNP ( B ′) vs the control group, P
    Anti Gst Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gst antibody/product/Millipore
    Average 99 stars, based on 11 article reviews
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    anti gst antibody - by Bioz Stars, 2020-09
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    92
    Millipore btx 1 bsa
    Construction and characterization of colloidal gold strip test; ( A ) the description of strip test results. In the absence of <t>BTX-1</t> in the sample solution, both two line exits on the control and test zone, indicating negative. Only one control line stands positive for enough toxin binding to the <t>anti-BTX-1-BSA</t> McAb. If no lines, or only the test line was red, it indicated invalid results; ( B ) cross-reactivity of the test strip with other toxins, such as OA, DA, BTX-2, BTX-3, TTX, CTX, STX; ( C ) the detection limit of colloidal gold strip test for BTX-1; ( D ) the real sample solution detection of colloidal gold strip for BTX-1.
    Btx 1 Bsa, supplied by Millipore, 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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    btx 1 bsa - by Bioz Stars, 2020-09
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    98
    Millipore biotin streptavidin complexes
    Localization of injected <t>bNTF2-streptavidin</t> complexes. bNTF2 (A) or biotinylated BSA (B) was preincubated with FIT C-streptavidin and coinjected with TRITC-dextran as an injection marker into the cytoplasm or nucleus of BHK21 cells. Cells were incubated at 37°C for 1 h after injection and fixed, and fluorescent proteins were visualized directly.
    Biotin Streptavidin Complexes, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    CPP-MT and CPP-MT-AuNP translocate intact human sperm. Notes: Sperm were incubated with 20 µg/mL of CPP-MT ( A ), CPP-MT-AuNP ( B ), or non-permeable GST-Rab3A ( C , control) for 3 hours at 37°C in HTF medium. Afterward, cells were treated with trypsin to eliminate external probe. Then, cells were fixed and spotted on glass slides and labeled with Cy3-coupled anti-GST antibody. At least 100 cells were quantified, and the population of sperm showing positive label was recorded. The bars represent mean values ± SEM, calculated from three independent experiments. The images are representative of the experimental conditions described. One-way ANOVA shows statistically significant difference between relative labeled cells percentages incubated with CPP-MT ( A ′) or CPP-MT-AuNP ( B ′) vs the control group, P

    Journal: International Journal of Nanomedicine

    Article Title: A TEM-traceable physiologically functional gold nanoprobe that permeates non-endocytic cells

    doi: 10.2147/IJN.S168149

    Figure Lengend Snippet: CPP-MT and CPP-MT-AuNP translocate intact human sperm. Notes: Sperm were incubated with 20 µg/mL of CPP-MT ( A ), CPP-MT-AuNP ( B ), or non-permeable GST-Rab3A ( C , control) for 3 hours at 37°C in HTF medium. Afterward, cells were treated with trypsin to eliminate external probe. Then, cells were fixed and spotted on glass slides and labeled with Cy3-coupled anti-GST antibody. At least 100 cells were quantified, and the population of sperm showing positive label was recorded. The bars represent mean values ± SEM, calculated from three independent experiments. The images are representative of the experimental conditions described. One-way ANOVA shows statistically significant difference between relative labeled cells percentages incubated with CPP-MT ( A ′) or CPP-MT-AuNP ( B ′) vs the control group, P

    Article Snippet: The sperms were labeled with 20 µg/mL anti-GST antibody (overnight at 4°C in 1% BSA-PBS), followed by Cy3-labeled anti-rabbit IgG as secondary antibodies (4 µg/mL in 1% BSA) for 1 hour at 4°C.

    Techniques: Conditioned Place Preference, Incubation, Labeling

    Construction and characterization of colloidal gold strip test; ( A ) the description of strip test results. In the absence of BTX-1 in the sample solution, both two line exits on the control and test zone, indicating negative. Only one control line stands positive for enough toxin binding to the anti-BTX-1-BSA McAb. If no lines, or only the test line was red, it indicated invalid results; ( B ) cross-reactivity of the test strip with other toxins, such as OA, DA, BTX-2, BTX-3, TTX, CTX, STX; ( C ) the detection limit of colloidal gold strip test for BTX-1; ( D ) the real sample solution detection of colloidal gold strip for BTX-1.

    Journal: Toxins

    Article Title: Preparation of Monoclonal Antibody for Brevetoxin 1 and Development of Ic-ELISA and Colloidal Gold Strip to Detect Brevetoxin 1

    doi: 10.3390/toxins10020075

    Figure Lengend Snippet: Construction and characterization of colloidal gold strip test; ( A ) the description of strip test results. In the absence of BTX-1 in the sample solution, both two line exits on the control and test zone, indicating negative. Only one control line stands positive for enough toxin binding to the anti-BTX-1-BSA McAb. If no lines, or only the test line was red, it indicated invalid results; ( B ) cross-reactivity of the test strip with other toxins, such as OA, DA, BTX-2, BTX-3, TTX, CTX, STX; ( C ) the detection limit of colloidal gold strip test for BTX-1; ( D ) the real sample solution detection of colloidal gold strip for BTX-1.

    Article Snippet: The reason was that BTX-1 will bind with antibody-gold nanoparticle conjugates, so it makes the antibody-gold nanoparticle conjugates fail to bind with the BTX-1-BSA that was coated onto the Millipore 135 NC membrane (Jieyi biotech Co., shanghai, china).

    Techniques: Stripping Membranes, Binding Assay

    Identification of the conjugates and anti-serum titer. ( A , B ) Analysis of conjugates and carrier protein by non-denaturing agarose electrophoris. ( A ) Lane 1: BTX-1-BSA conjugates sample, Lane 2: BSA; ( B ) Lane 1: BTX-1-OVA conjugates sample, Lane 2: OVA. ( C ) Titer of mice anti-serum measured by indirect ELISA. Mouse 1 and 2 were immunized with BTX-1-OVA conjugate, and the control was immunized with only adjuvant and PBS.

    Journal: Toxins

    Article Title: Preparation of Monoclonal Antibody for Brevetoxin 1 and Development of Ic-ELISA and Colloidal Gold Strip to Detect Brevetoxin 1

    doi: 10.3390/toxins10020075

    Figure Lengend Snippet: Identification of the conjugates and anti-serum titer. ( A , B ) Analysis of conjugates and carrier protein by non-denaturing agarose electrophoris. ( A ) Lane 1: BTX-1-BSA conjugates sample, Lane 2: BSA; ( B ) Lane 1: BTX-1-OVA conjugates sample, Lane 2: OVA. ( C ) Titer of mice anti-serum measured by indirect ELISA. Mouse 1 and 2 were immunized with BTX-1-OVA conjugate, and the control was immunized with only adjuvant and PBS.

    Article Snippet: The reason was that BTX-1 will bind with antibody-gold nanoparticle conjugates, so it makes the antibody-gold nanoparticle conjugates fail to bind with the BTX-1-BSA that was coated onto the Millipore 135 NC membrane (Jieyi biotech Co., shanghai, china).

    Techniques: Mouse Assay, Indirect ELISA

    Localization of injected bNTF2-streptavidin complexes. bNTF2 (A) or biotinylated BSA (B) was preincubated with FIT C-streptavidin and coinjected with TRITC-dextran as an injection marker into the cytoplasm or nucleus of BHK21 cells. Cells were incubated at 37°C for 1 h after injection and fixed, and fluorescent proteins were visualized directly.

    Journal: Molecular Biology of the Cell

    Article Title: Monoclonal Antibodies to NTF2 Inhibit Nuclear Protein Import by Preventing Nuclear Translocation of the GTPase Ran

    doi:

    Figure Lengend Snippet: Localization of injected bNTF2-streptavidin complexes. bNTF2 (A) or biotinylated BSA (B) was preincubated with FIT C-streptavidin and coinjected with TRITC-dextran as an injection marker into the cytoplasm or nucleus of BHK21 cells. Cells were incubated at 37°C for 1 h after injection and fixed, and fluorescent proteins were visualized directly.

    Article Snippet: For preparation of biotin-streptavidin complexes, an excess of biotinylated protein was mixed with FITC-neutravidin on an end-over-end rotator at 4°C for several hours and then filtered for microinjection using 0.22-μm Millipore (Bedford, MA) centrifugal filtration units.

    Techniques: Injection, Marker, Incubation