succinimidyl ester  (Thermo Fisher)


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    Structured Review

    Thermo Fisher succinimidyl ester
    Cell fusion of MSCs and NP cells during direct co-culture. A–C: An exemplar flow cytometry analysis for CFDA labeled MSCs and SNARF labeled NP cells after 7 days: A) CFDA labeled MSCs alone; B) SNARF labeled NP cells alone; C) Co-culture of CFDA labeled MSCs and SNARF labeled NP cells. D–F: Fluorescence microscopy of sorted double labeled cells from region R4: D) Cells labeled with CFDA; E) Cells labeled with SNARF; F) Color combine; enlargement of a 100× magnification. G) Percentage of cell fusion after 1, 3 and 7 days calculated from flow cytometry data. Error bars indicate standard error of the mean. Abbreviations: MSC: mesenchymal stem cell; NP: nucleus pulposus; CFDA: 5,6 caboxyfluorescein diactetae, <t>succinimidyl</t> ester; SNARF: carboxylic acid, acetate, SE.
    Succinimidyl Ester, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/succinimidyl ester/product/Thermo Fisher
    Average 99 stars, based on 31 article reviews
    Price from $9.99 to $1999.99
    succinimidyl ester - by Bioz Stars, 2020-01
    99/100 stars

    Images

    1) Product Images from "Bi-Directional Exchange of Membrane Components Occurs during Co-Culture of Mesenchymal Stem Cells and Nucleus Pulposus Cells"

    Article Title: Bi-Directional Exchange of Membrane Components Occurs during Co-Culture of Mesenchymal Stem Cells and Nucleus Pulposus Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0033739

    Cell fusion of MSCs and NP cells during direct co-culture. A–C: An exemplar flow cytometry analysis for CFDA labeled MSCs and SNARF labeled NP cells after 7 days: A) CFDA labeled MSCs alone; B) SNARF labeled NP cells alone; C) Co-culture of CFDA labeled MSCs and SNARF labeled NP cells. D–F: Fluorescence microscopy of sorted double labeled cells from region R4: D) Cells labeled with CFDA; E) Cells labeled with SNARF; F) Color combine; enlargement of a 100× magnification. G) Percentage of cell fusion after 1, 3 and 7 days calculated from flow cytometry data. Error bars indicate standard error of the mean. Abbreviations: MSC: mesenchymal stem cell; NP: nucleus pulposus; CFDA: 5,6 caboxyfluorescein diactetae, succinimidyl ester; SNARF: carboxylic acid, acetate, SE.
    Figure Legend Snippet: Cell fusion of MSCs and NP cells during direct co-culture. A–C: An exemplar flow cytometry analysis for CFDA labeled MSCs and SNARF labeled NP cells after 7 days: A) CFDA labeled MSCs alone; B) SNARF labeled NP cells alone; C) Co-culture of CFDA labeled MSCs and SNARF labeled NP cells. D–F: Fluorescence microscopy of sorted double labeled cells from region R4: D) Cells labeled with CFDA; E) Cells labeled with SNARF; F) Color combine; enlargement of a 100× magnification. G) Percentage of cell fusion after 1, 3 and 7 days calculated from flow cytometry data. Error bars indicate standard error of the mean. Abbreviations: MSC: mesenchymal stem cell; NP: nucleus pulposus; CFDA: 5,6 caboxyfluorescein diactetae, succinimidyl ester; SNARF: carboxylic acid, acetate, SE.

    Techniques Used: Co-Culture Assay, Flow Cytometry, Cytometry, Labeling, Fluorescence, Microscopy

    Transfer of membrane components during direct co-culture of MSC and NP cells. A–C: Exemplar flow cytometry to quantify transfer of membrane (DiL) components after 7 days. A) Dot plots for unlabeled MSCs and NP cells. B) Dot plots for CFDA and DiL labeled MSCs and NP cells. C) Dot plots for CFDA and DiL double labeled MSCs co-cultured with unlabeled NP cells; CFDA and DiL double labeled NP cells co-cultured with unlabeled MSCs. D) Percentages of DiL transfer after 1, 3 and 7 days from a labeled cell to an unlabeled cell during direct co-culture calculated from flow cytometry data. Error bars indicate standard error of the mean. Abbreviations: MSC: mesenchymal stem cell; NP: nucleus pulposus; CFDA: 5,6 carboxyfluorescein diactetae, succinimidyl ester; DiL: Vybrant CM-DiL cell-labeling solution.
    Figure Legend Snippet: Transfer of membrane components during direct co-culture of MSC and NP cells. A–C: Exemplar flow cytometry to quantify transfer of membrane (DiL) components after 7 days. A) Dot plots for unlabeled MSCs and NP cells. B) Dot plots for CFDA and DiL labeled MSCs and NP cells. C) Dot plots for CFDA and DiL double labeled MSCs co-cultured with unlabeled NP cells; CFDA and DiL double labeled NP cells co-cultured with unlabeled MSCs. D) Percentages of DiL transfer after 1, 3 and 7 days from a labeled cell to an unlabeled cell during direct co-culture calculated from flow cytometry data. Error bars indicate standard error of the mean. Abbreviations: MSC: mesenchymal stem cell; NP: nucleus pulposus; CFDA: 5,6 carboxyfluorescein diactetae, succinimidyl ester; DiL: Vybrant CM-DiL cell-labeling solution.

    Techniques Used: Co-Culture Assay, Flow Cytometry, Cytometry, Labeling, Cell Culture

    Incorporation of DiL-labeled microvesicles into MSC and NP cells during direct co-culture. Exemplar flow cytometry analysis. A) MVs derived from a direct co-culture of MSCs and NP cells (one cell population was DiL labeled) and incubated with the unlabeled cell population. B) MV-free supernatant after ultracentrifugation. C) Conditioned medium derived from a direct co-culture of MSCs and NP cells (one cell population was DiL labeled). Abbreviations: MSC: mesenchymal stem cell; NP: nucleus pulposus; MV: microvesicles; SN: supernatant; CM: conditioned medium; CFDA: 5,6 caboxyfluorescein diactetae, succinimidyl ester; DiL: Vybrant CM-DiL cell-labeling solution.
    Figure Legend Snippet: Incorporation of DiL-labeled microvesicles into MSC and NP cells during direct co-culture. Exemplar flow cytometry analysis. A) MVs derived from a direct co-culture of MSCs and NP cells (one cell population was DiL labeled) and incubated with the unlabeled cell population. B) MV-free supernatant after ultracentrifugation. C) Conditioned medium derived from a direct co-culture of MSCs and NP cells (one cell population was DiL labeled). Abbreviations: MSC: mesenchymal stem cell; NP: nucleus pulposus; MV: microvesicles; SN: supernatant; CM: conditioned medium; CFDA: 5,6 caboxyfluorescein diactetae, succinimidyl ester; DiL: Vybrant CM-DiL cell-labeling solution.

    Techniques Used: Labeling, Co-Culture Assay, Flow Cytometry, Cytometry, Derivative Assay, Incubation

    2) Product Images from "Hemagglutinin-based polyanhydride nanovaccines against H5N1 influenza elicit protective virus neutralizing titers and cell-mediated immunity"

    Article Title: Hemagglutinin-based polyanhydride nanovaccines against H5N1 influenza elicit protective virus neutralizing titers and cell-mediated immunity

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S72264

    Enhanced CD4 + T cell memory with poly I:C polyanhydride nanovaccine. Notes: Draining lymph nodes were harvested from prime/boost/boost immunized mice 63 days after the primary immunization. Ex vivo antigen stimulation and CFSE-labeling demonstrated enhanced CD4 + T cell proliferation of mice immunized with poly I:C nanovaccines. Error bars represent the standard error of the mean. Different letters indicate statistical significance among treatments. P ≤0.0147. Abbreviations: CFSE, 5-(and-6)-carboxyfluorescein diacetate, succinimidyl ester; MPLA, monophosphoryl lipid A; sH5 3 , soluble H5 hemagglutinin trimer; poly I:C, polyinosinic-polycytidylic acid..
    Figure Legend Snippet: Enhanced CD4 + T cell memory with poly I:C polyanhydride nanovaccine. Notes: Draining lymph nodes were harvested from prime/boost/boost immunized mice 63 days after the primary immunization. Ex vivo antigen stimulation and CFSE-labeling demonstrated enhanced CD4 + T cell proliferation of mice immunized with poly I:C nanovaccines. Error bars represent the standard error of the mean. Different letters indicate statistical significance among treatments. P ≤0.0147. Abbreviations: CFSE, 5-(and-6)-carboxyfluorescein diacetate, succinimidyl ester; MPLA, monophosphoryl lipid A; sH5 3 , soluble H5 hemagglutinin trimer; poly I:C, polyinosinic-polycytidylic acid..

    Techniques Used: Mouse Assay, Ex Vivo, Labeling

    3) Product Images from "The focal adhesion kinase inhibitor PF-562,271 impairs primary CD4+ T cell activation"

    Article Title: The focal adhesion kinase inhibitor PF-562,271 impairs primary CD4+ T cell activation

    Journal: Biochemical pharmacology

    doi: 10.1016/j.bcp.2013.07.024

    PF-562,271 inhibits T cell proliferation. OVA peptide expanded mouse CD4+ cells were stained with 0.25 μM carboxyfluorescein diacetate, succinimidyl ester and stimulated with anti-CD3/CD28 coated beads (A), irradiated splenocytes loaded 2.5 μg/mL
    Figure Legend Snippet: PF-562,271 inhibits T cell proliferation. OVA peptide expanded mouse CD4+ cells were stained with 0.25 μM carboxyfluorescein diacetate, succinimidyl ester and stimulated with anti-CD3/CD28 coated beads (A), irradiated splenocytes loaded 2.5 μg/mL

    Techniques Used: Staining, Irradiation

    4) Product Images from "Multiple roles of filopodial dynamics in particle capture and phagocytosis and phenotypes of Cdc42 and Myo10 deletion"

    Article Title: Multiple roles of filopodial dynamics in particle capture and phagocytosis and phenotypes of Cdc42 and Myo10 deletion

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M116.766923

    Dectin-1 mediates zymosan binding and internalization. A , fluorescence emission spectra of pHrodo-conjugated zymosan particles suspended in buffers with different pH values, ranging from pH 4 to 8. Particles were labeled using the succinimidyl ester of pHrodo. B , top panel : DIC and fluorescence images of pH-sensitive pHrodo-conjugated zymosan particles superfused with media buffered to different pH values, as indicated. Bottom panel : time-lapse images showing the ingestion and acidification of pHrodo-conjugated zymosan particles. C , DIC image, with superimposed red fluorescence, of two wild-type (WT) macrophages, which have ingested multiple pHrodo-conjugated zymosan particles. Red fluorescence indicates acidification of phagosomes. Scale bar , 10 μm. D , box plots of the number of attached ( left ) and red fluorescent ( right ) pHrodo-conjugated zymosan particles in WT, Toll-like receptor 2/4 double knock-out (TLR2/4 dKO), mannose receptor knock-out ( MR −/− ), and Dectin-1 knock-out ( Dectin-1 −/− ) macrophages. Cells, seeded in fibronectin-coated μ-slide I chambers, were incubated with particles for 15 min before washing. *, p
    Figure Legend Snippet: Dectin-1 mediates zymosan binding and internalization. A , fluorescence emission spectra of pHrodo-conjugated zymosan particles suspended in buffers with different pH values, ranging from pH 4 to 8. Particles were labeled using the succinimidyl ester of pHrodo. B , top panel : DIC and fluorescence images of pH-sensitive pHrodo-conjugated zymosan particles superfused with media buffered to different pH values, as indicated. Bottom panel : time-lapse images showing the ingestion and acidification of pHrodo-conjugated zymosan particles. C , DIC image, with superimposed red fluorescence, of two wild-type (WT) macrophages, which have ingested multiple pHrodo-conjugated zymosan particles. Red fluorescence indicates acidification of phagosomes. Scale bar , 10 μm. D , box plots of the number of attached ( left ) and red fluorescent ( right ) pHrodo-conjugated zymosan particles in WT, Toll-like receptor 2/4 double knock-out (TLR2/4 dKO), mannose receptor knock-out ( MR −/− ), and Dectin-1 knock-out ( Dectin-1 −/− ) macrophages. Cells, seeded in fibronectin-coated μ-slide I chambers, were incubated with particles for 15 min before washing. *, p

    Techniques Used: Binding Assay, Fluorescence, Labeling, Knock-Out, Incubation

    5) Product Images from "A Role for the Eph Ligand Ephrin-A3 in Entorhino-Hippocampal Axon Targeting"

    Article Title: A Role for the Eph Ligand Ephrin-A3 in Entorhino-Hippocampal Axon Targeting

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.19-20-08885.1999

    Inhibition of neurite outgrowth from entorhinal explants by ephrin-A3. Entorhinal explants were cultivated on a confluent membrane layer obtained from 3T3 mock-transfected ( n = 41) ( A ), ephrin-A3- ( n = 62) ( C ), and ephrin-A5- ( n = 31) ( E ) transfected cells. Neurites were visualized with a fluorescent vital dye, 5- and 6-carboxyfluorescein diacetate, succinimidyl ester. D , PI-PLC treatment of the obtained membranes ( n = 36) significantly prevents ephrin-A3 inhibition of entorhinal neurite outgrowth, whereas the effect of PI-PLC treatment on neurite outgrowth inhibition ( n = 25) was not significant in the case of ephrin-A5 ( F ). B , Quantification of entorhinal neurite length under the different conditions (±SEM). The differences of entorhinal outgrowth length were significant between ephrin-A3 treatment and all other experimental conditions (* p
    Figure Legend Snippet: Inhibition of neurite outgrowth from entorhinal explants by ephrin-A3. Entorhinal explants were cultivated on a confluent membrane layer obtained from 3T3 mock-transfected ( n = 41) ( A ), ephrin-A3- ( n = 62) ( C ), and ephrin-A5- ( n = 31) ( E ) transfected cells. Neurites were visualized with a fluorescent vital dye, 5- and 6-carboxyfluorescein diacetate, succinimidyl ester. D , PI-PLC treatment of the obtained membranes ( n = 36) significantly prevents ephrin-A3 inhibition of entorhinal neurite outgrowth, whereas the effect of PI-PLC treatment on neurite outgrowth inhibition ( n = 25) was not significant in the case of ephrin-A5 ( F ). B , Quantification of entorhinal neurite length under the different conditions (±SEM). The differences of entorhinal outgrowth length were significant between ephrin-A3 treatment and all other experimental conditions (* p

    Techniques Used: Inhibition, Transfection, Planar Chromatography

    Related Articles

    Synthesized:

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    Cytometry:

    Article Title: Bi-Directional Exchange of Membrane Components Occurs during Co-Culture of Mesenchymal Stem Cells and Nucleus Pulposus Cells
    Article Snippet: Assessment of cell fusion Cellular fusion was ascertained by 5,6 carboxyfluorescein diactetate, succinimidyl ester (CFDA; Invitrogen) and SNARF-1 carboxylic acid, acetate, succinimidyl ester (SNARF-1; (Invitrogen). .. After 1, 3 and 7 days, all co-cultures and controls were analysed for CFDA and SNARF-1 fluorescence using flow cytometry.

    Article Title: Apolipoprotein A-I gene transfer exerts immunomodulatory effects and reduces vascular inflammation and fibrosis in ob/ob mice
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    Co-Culture Assay:

    Article Title: Bi-Directional Exchange of Membrane Components Occurs during Co-Culture of Mesenchymal Stem Cells and Nucleus Pulposus Cells
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    Incubation:

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    Article Title: The Mammalian Septin MSF Localizes with Microtubules and Is Required for Completion of Cytokinesis
    Article Snippet: .. Approximately 60 mg of either anti-MSF or anti-Nedd5 IgG was incubated with 6-(tetramethylrhodamine-5-(and-6)-carboxamido)hexanoic acid, succinimidyl ester, or 6-(fluorescein-5-(and-6)-carboxamido)hexanoic acid, succinimidyl ester (Molecular Probes), respectively, in a 1:10 M ratio (protein:dye). .. Labeling was performed according to the manufacturer's directions with the unlabeled conjugate removed from the reaction by column chromatography over a PD-10 column (Amersham).

    Article Title: Glycan-mediated enhancement of reovirus receptor binding
    Article Snippet: .. For fluorescent labeling, reovirus particles were diluted into fresh 50 mM sodium bicarbonate (pH 8.5; 6 × 1012 particles/mL) and incubated with 20 μM succinimidyl ester of Alexa Flour 488 (Invitrogen) at room temperature for 90 min in the dark . ..

    Article Title: Multiple roles of filopodial dynamics in particle capture and phagocytosis and phenotypes of Cdc42 and Myo10 deletion
    Article Snippet: The following BioParticles were used: Alexa Fluor 594-conjugated E. coli (K-12 strain) particles (E23370; Thermo Fisher Scientific), Alexa Fluor 594-conjugated zymosan (polysaccharide prepared from the cell wall of S. cerevisiae (bakers' yeast), catalogue number ; Thermo Fisher Scientific) and pHrodo-conjugated zymosan, prepared using unlabeled zymosan (Z2849; Thermo Fisher Scientific) and the succinimidyl ester of pHrodo red ( ; Thermo Fisher Scientific), which has absorbance and emission maxima of 560 and 585 nm, respectively. .. The 0.5% (w/v) stock solution was diluted 1:10 before use, and the beads were rendered red fluorescent by incubation with Alexa Fluor 594-conjugated goat anti-mouse IgG antibodies (ab150116; Abcam), followed by a wash step.

    Article Title: Sulfiredoxin redox-sensitive interaction with S100A4 and non-muscle myosin IIA regulates cancer cell motility
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    Article Title: Hemagglutinin-based polyanhydride nanovaccines against H5N1 influenza elicit protective virus neutralizing titers and cell-mediated immunity
    Article Snippet: Single cell populations were labeled with 2.5 μM 5-(and-6)- carboxyfluorescein diacetate, succinimidyl ester (5[6]-CFDA, SE; CFSE; Thermo Fisher Scientific, Waltham, MA, USA). .. Cells (2.5×105 ) were incubated in 96-well U-bottom plates with 0.5 μg of sH53 antigen for 96 hours.

    Stripping Membranes:

    Article Title: A Role for the Eph Ligand Ephrin-A3 in Entorhino-Hippocampal Axon Targeting
    Article Snippet: Paragraph title: Explant outgrowth and stripe assay ... Neurites growing out from the explants were visualized with a fluorescent vital dye, 5 and 6-carboxyfluorescein diacetate, succinimidyl ester (Molecular Probes), which labels all living cells and their processes, or by immunolabeling with a neurofilament antibody (Boehringer Mannheim).

    Expressing:

    Article Title: Hemagglutinin-based polyanhydride nanovaccines against H5N1 influenza elicit protective virus neutralizing titers and cell-mediated immunity
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    Modification:

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    Conjugation Assay:

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    Transfection:

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    Concentration Assay:

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    Article Title: Glycan-mediated enhancement of reovirus receptor binding
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    Protease Inhibitor:

    Article Title: The focal adhesion kinase inhibitor PF-562,271 impairs primary CD4+ T cell activation
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    Immunolabeling:

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    Article Snippet: .. Neurites growing out from the explants were visualized with a fluorescent vital dye, 5 and 6-carboxyfluorescein diacetate, succinimidyl ester (Molecular Probes), which labels all living cells and their processes, or by immunolabeling with a neurofilament antibody (Boehringer Mannheim). ..

    Cell Culture:

    Article Title: Apolipoprotein A-I gene transfer exerts immunomodulatory effects and reduces vascular inflammation and fibrosis in ob/ob mice
    Article Snippet: (Splenocyte) mononuclear cell, CD4-, and CD8-T cell proliferation Splenocytes were labeled with 10 μM of succinimidyl ester of carboxyfluorescein diacetate (CFSE Cell Tract™; Invitrogen, Carlbard, CA, USA) to be able to measure cell proliferation, indicative for MNC activation, as described previously [ ]. .. Therefore, splenocytes were cultured in RPMI1640 medium (Invitrogen, Heidelberg, Germany), supplemented with 10 % FBS and 1 % penicillin/streptomycin for 72 h. Then, cells were stained with monoclonal anti-CD4 or anti-CD8 antibodies (BD Biosciences, Franklin Lakes, NJ, USA), followed by flow cytometry on a MACSQuant Analyzer (Miltenyi Biotec, Bergisch Gladbach, Germany) and analysis with FlowJo 8.7. software (Tree Star) for the calculation of the division index, i.e. the average number of cell divisions that the responding cells undergo (i.e., ignores peak 0).

    Imaging:

    Article Title: Multiple roles of filopodial dynamics in particle capture and phagocytosis and phenotypes of Cdc42 and Myo10 deletion
    Article Snippet: The suspension of BioParticles in live cell imaging medium was sonicated to disperse the particles, and 100 μl was pipetted into a μ-slide I chamber before starting time-lapse imaging via spinning disk confocal microscopy. .. The following BioParticles were used: Alexa Fluor 594-conjugated E. coli (K-12 strain) particles (E23370; Thermo Fisher Scientific), Alexa Fluor 594-conjugated zymosan (polysaccharide prepared from the cell wall of S. cerevisiae (bakers' yeast), catalogue number ; Thermo Fisher Scientific) and pHrodo-conjugated zymosan, prepared using unlabeled zymosan (Z2849; Thermo Fisher Scientific) and the succinimidyl ester of pHrodo red ( ; Thermo Fisher Scientific), which has absorbance and emission maxima of 560 and 585 nm, respectively.

    Sonication:

    Article Title: Multiple roles of filopodial dynamics in particle capture and phagocytosis and phenotypes of Cdc42 and Myo10 deletion
    Article Snippet: The suspension of BioParticles in live cell imaging medium was sonicated to disperse the particles, and 100 μl was pipetted into a μ-slide I chamber before starting time-lapse imaging via spinning disk confocal microscopy. .. The following BioParticles were used: Alexa Fluor 594-conjugated E. coli (K-12 strain) particles (E23370; Thermo Fisher Scientific), Alexa Fluor 594-conjugated zymosan (polysaccharide prepared from the cell wall of S. cerevisiae (bakers' yeast), catalogue number ; Thermo Fisher Scientific) and pHrodo-conjugated zymosan, prepared using unlabeled zymosan (Z2849; Thermo Fisher Scientific) and the succinimidyl ester of pHrodo red ( ; Thermo Fisher Scientific), which has absorbance and emission maxima of 560 and 585 nm, respectively.

    Binding Assay:

    Article Title: Hemagglutinin-based polyanhydride nanovaccines against H5N1 influenza elicit protective virus neutralizing titers and cell-mediated immunity
    Article Snippet: Single cell populations were labeled with 2.5 μM 5-(and-6)- carboxyfluorescein diacetate, succinimidyl ester (5[6]-CFDA, SE; CFSE; Thermo Fisher Scientific, Waltham, MA, USA). .. Cell suspensions were blocked for nonspecific antibody binding using 0.1 mg/mL Rat IgG (Sigma-Aldrich Co.) and 10 μg/mL mouse anti-CD16/32 (eBioscience, San Diego, CA, USA).

    Immunofluorescence:

    Article Title: Cell-cycle-regulated activation of Akt kinase by phosphorylation at its carboxyl terminus
    Article Snippet: Paragraph title: Fixing and staining cells for immunofluorescence ... DNA was stained by incubating in 10 μM 4′,6-diamidino-2-phenylindole (DAPI) for 5 min. To label protein mass, fixed, permeabilized samples were incubated with 0.04 μg ml−1 succinimidyl ester linked dye diluted in PBS (Alexa Fluor 647 carboxylic acid, succinimidyl ester; Invitrogen, A-20106).

    Plaque Assay:

    Article Title: Glycan-mediated enhancement of reovirus receptor binding
    Article Snippet: Viral titers were determined by plaque assay using L929 cells . .. For fluorescent labeling, reovirus particles were diluted into fresh 50 mM sodium bicarbonate (pH 8.5; 6 × 1012 particles/mL) and incubated with 20 μM succinimidyl ester of Alexa Flour 488 (Invitrogen) at room temperature for 90 min in the dark .

    Fluorescence:

    Article Title: Bi-Directional Exchange of Membrane Components Occurs during Co-Culture of Mesenchymal Stem Cells and Nucleus Pulposus Cells
    Article Snippet: Assessment of cell fusion Cellular fusion was ascertained by 5,6 carboxyfluorescein diactetate, succinimidyl ester (CFDA; Invitrogen) and SNARF-1 carboxylic acid, acetate, succinimidyl ester (SNARF-1; (Invitrogen). .. After 1, 3 and 7 days, all co-cultures and controls were analysed for CFDA and SNARF-1 fluorescence using flow cytometry.

    Mutagenesis:

    Article Title: Sulfiredoxin redox-sensitive interaction with S100A4 and non-muscle myosin IIA regulates cancer cell motility
    Article Snippet: Purified Srx (C99S) protein was incubated with 5x excess of succinimidyl ester of Alexa® 546 Fluor carboxylic acid (Invitrogen) and S100A4 (S100A4-SSG) purified protein was incubated with 5x excess of succinimidyl ester of QSY®35 acetic acid (Invitrogen) at room temperature, overnight, under constant agitation to label primary amines according to manufacturer’s recommendations. .. Spectroscopic analysis of Alexa® 546 Fluor label incorporation indicated ~2.0 mol of dye per 1 mol of Srx or C99S mutant monomer.

    Flow Cytometry:

    Article Title: Bi-Directional Exchange of Membrane Components Occurs during Co-Culture of Mesenchymal Stem Cells and Nucleus Pulposus Cells
    Article Snippet: Assessment of cell fusion Cellular fusion was ascertained by 5,6 carboxyfluorescein diactetate, succinimidyl ester (CFDA; Invitrogen) and SNARF-1 carboxylic acid, acetate, succinimidyl ester (SNARF-1; (Invitrogen). .. After 1, 3 and 7 days, all co-cultures and controls were analysed for CFDA and SNARF-1 fluorescence using flow cytometry.

    Article Title: Apolipoprotein A-I gene transfer exerts immunomodulatory effects and reduces vascular inflammation and fibrosis in ob/ob mice
    Article Snippet: (Splenocyte) mononuclear cell, CD4-, and CD8-T cell proliferation Splenocytes were labeled with 10 μM of succinimidyl ester of carboxyfluorescein diacetate (CFSE Cell Tract™; Invitrogen, Carlbard, CA, USA) to be able to measure cell proliferation, indicative for MNC activation, as described previously [ ]. .. Therefore, splenocytes were cultured in RPMI1640 medium (Invitrogen, Heidelberg, Germany), supplemented with 10 % FBS and 1 % penicillin/streptomycin for 72 h. Then, cells were stained with monoclonal anti-CD4 or anti-CD8 antibodies (BD Biosciences, Franklin Lakes, NJ, USA), followed by flow cytometry on a MACSQuant Analyzer (Miltenyi Biotec, Bergisch Gladbach, Germany) and analysis with FlowJo 8.7. software (Tree Star) for the calculation of the division index, i.e. the average number of cell divisions that the responding cells undergo (i.e., ignores peak 0).

    Article Title: Hemagglutinin-based polyanhydride nanovaccines against H5N1 influenza elicit protective virus neutralizing titers and cell-mediated immunity
    Article Snippet: Paragraph title: Flow cytometry for T cell memory populations ... Single cell populations were labeled with 2.5 μM 5-(and-6)- carboxyfluorescein diacetate, succinimidyl ester (5[6]-CFDA, SE; CFSE; Thermo Fisher Scientific, Waltham, MA, USA).

    Microscopy:

    Article Title: A Role for the Eph Ligand Ephrin-A3 in Entorhino-Hippocampal Axon Targeting
    Article Snippet: Neurites growing out from the explants were visualized with a fluorescent vital dye, 5 and 6-carboxyfluorescein diacetate, succinimidyl ester (Molecular Probes), which labels all living cells and their processes, or by immunolabeling with a neurofilament antibody (Boehringer Mannheim). .. Neurite growth from the explant was examined and photographed with FITC optics on an epifluorescence microscope.

    Size-exclusion Chromatography:

    Article Title: Cancer mutations of the tumor suppressor SPOP disrupt the formation of active, phase-separated compartments
    Article Snippet: GST-SPOPMATH was expressed in BL21 RIPL cells in LB medium, and purified as previously described by Glutathione-sepharose, TEV cleavage during dialysis, ion-exchange by SP column, and SEC ( ). .. Small fractions of each construct were labeled with 4X Oregon Green 488 Carboxylic Acid, Succinimidyl Ester, 5 isomer (Cat #O6147 Thermo Fisher) at 4 ºC overnight.

    Protein Purification:

    Article Title: Cancer mutations of the tumor suppressor SPOP disrupt the formation of active, phase-separated compartments
    Article Snippet: Paragraph title: Protein Purification ... Small fractions of each construct were labeled with 4X Oregon Green 488 Carboxylic Acid, Succinimidyl Ester, 5 isomer (Cat #O6147 Thermo Fisher) at 4 ºC overnight.

    Opsonophagocytic Assay:

    Article Title: Evaluation of Multiplex Flow Cytometric Opsonophagocytic Assays for Determination of Functional Anticapsular Antibodies to Streptococcus pneumoniae
    Article Snippet: S. pneumoniae serotypes 1, 4, 6B, 9V, 14, 18C, 19F, and 23F were grown as previously described for the single-color-bacterium-based opsonophagocytic assay ( ). .. The bacterium-staining protocol was modified to use 1.25 mg/ml (5,6)-carboxytetramethylrhodamine, succinimidyl ester, (5,6)-TAMRA (6-carboxytetramethylrhodamine), succinimidyl ester (Molecular Probes, Eugene, Oreg.), or 0.25 mg/ml Texas Red-X-succinimidyl ester (Molecular Probes, Eugene, Oreg.) as an alternative to the 10 mg/ml 5,6 carboxyfluorescein used in the original protocol.

    Labeling:

    Article Title: Bi-Directional Exchange of Membrane Components Occurs during Co-Culture of Mesenchymal Stem Cells and Nucleus Pulposus Cells
    Article Snippet: Assessment of cell fusion Cellular fusion was ascertained by 5,6 carboxyfluorescein diactetate, succinimidyl ester (CFDA; Invitrogen) and SNARF-1 carboxylic acid, acetate, succinimidyl ester (SNARF-1; (Invitrogen). .. Prior to direct co-culture, MSCs were fluorescently labeled with a final concentration of 10 µM CFDA and NP cells were labeled with 10 µM SNARF-1.

    Article Title: Cancer mutations of the tumor suppressor SPOP disrupt the formation of active, phase-separated compartments
    Article Snippet: .. Small fractions of each construct were labeled with 4X Oregon Green 488 Carboxylic Acid, Succinimidyl Ester, 5 isomer (Cat #O6147 Thermo Fisher) at 4 ºC overnight. .. Un-reacted label was removed from the proteins by PD-10 column (Thermo Fisher 45–000-67).

    Article Title: Evaluation of Multiplex Flow Cytometric Opsonophagocytic Assays for Determination of Functional Anticapsular Antibodies to Streptococcus pneumoniae
    Article Snippet: Paragraph title: Preparation of fluorescently labeled bacteria. ... The bacterium-staining protocol was modified to use 1.25 mg/ml (5,6)-carboxytetramethylrhodamine, succinimidyl ester, (5,6)-TAMRA (6-carboxytetramethylrhodamine), succinimidyl ester (Molecular Probes, Eugene, Oreg.), or 0.25 mg/ml Texas Red-X-succinimidyl ester (Molecular Probes, Eugene, Oreg.) as an alternative to the 10 mg/ml 5,6 carboxyfluorescein used in the original protocol.

    Article Title: The Mammalian Septin MSF Localizes with Microtubules and Is Required for Completion of Cytokinesis
    Article Snippet: Approximately 60 mg of either anti-MSF or anti-Nedd5 IgG was incubated with 6-(tetramethylrhodamine-5-(and-6)-carboxamido)hexanoic acid, succinimidyl ester, or 6-(fluorescein-5-(and-6)-carboxamido)hexanoic acid, succinimidyl ester (Molecular Probes), respectively, in a 1:10 M ratio (protein:dye). .. Labeling was performed according to the manufacturer's directions with the unlabeled conjugate removed from the reaction by column chromatography over a PD-10 column (Amersham).

    Article Title: Glycan-mediated enhancement of reovirus receptor binding
    Article Snippet: .. For fluorescent labeling, reovirus particles were diluted into fresh 50 mM sodium bicarbonate (pH 8.5; 6 × 1012 particles/mL) and incubated with 20 μM succinimidyl ester of Alexa Flour 488 (Invitrogen) at room temperature for 90 min in the dark . ..

    Article Title: Apolipoprotein A-I gene transfer exerts immunomodulatory effects and reduces vascular inflammation and fibrosis in ob/ob mice
    Article Snippet: .. (Splenocyte) mononuclear cell, CD4-, and CD8-T cell proliferation Splenocytes were labeled with 10 μM of succinimidyl ester of carboxyfluorescein diacetate (CFSE Cell Tract™; Invitrogen, Carlbard, CA, USA) to be able to measure cell proliferation, indicative for MNC activation, as described previously [ ]. .. Therefore, splenocytes were cultured in RPMI1640 medium (Invitrogen, Heidelberg, Germany), supplemented with 10 % FBS and 1 % penicillin/streptomycin for 72 h. Then, cells were stained with monoclonal anti-CD4 or anti-CD8 antibodies (BD Biosciences, Franklin Lakes, NJ, USA), followed by flow cytometry on a MACSQuant Analyzer (Miltenyi Biotec, Bergisch Gladbach, Germany) and analysis with FlowJo 8.7. software (Tree Star) for the calculation of the division index, i.e. the average number of cell divisions that the responding cells undergo (i.e., ignores peak 0).

    Article Title: Hemagglutinin-based polyanhydride nanovaccines against H5N1 influenza elicit protective virus neutralizing titers and cell-mediated immunity
    Article Snippet: .. Single cell populations were labeled with 2.5 μM 5-(and-6)- carboxyfluorescein diacetate, succinimidyl ester (5[6]-CFDA, SE; CFSE; Thermo Fisher Scientific, Waltham, MA, USA). .. Cells (2.5×105 ) were incubated in 96-well U-bottom plates with 0.5 μg of sH53 antigen for 96 hours.

    Confocal Microscopy:

    Article Title: Multiple roles of filopodial dynamics in particle capture and phagocytosis and phenotypes of Cdc42 and Myo10 deletion
    Article Snippet: The suspension of BioParticles in live cell imaging medium was sonicated to disperse the particles, and 100 μl was pipetted into a μ-slide I chamber before starting time-lapse imaging via spinning disk confocal microscopy. .. The following BioParticles were used: Alexa Fluor 594-conjugated E. coli (K-12 strain) particles (E23370; Thermo Fisher Scientific), Alexa Fluor 594-conjugated zymosan (polysaccharide prepared from the cell wall of S. cerevisiae (bakers' yeast), catalogue number ; Thermo Fisher Scientific) and pHrodo-conjugated zymosan, prepared using unlabeled zymosan (Z2849; Thermo Fisher Scientific) and the succinimidyl ester of pHrodo red ( ; Thermo Fisher Scientific), which has absorbance and emission maxima of 560 and 585 nm, respectively.

    Staining:

    Article Title: Cell-cycle-regulated activation of Akt kinase by phosphorylation at its carboxyl terminus
    Article Snippet: .. DNA was stained by incubating in 10 μM 4′,6-diamidino-2-phenylindole (DAPI) for 5 min. To label protein mass, fixed, permeabilized samples were incubated with 0.04 μg ml−1 succinimidyl ester linked dye diluted in PBS (Alexa Fluor 647 carboxylic acid, succinimidyl ester; Invitrogen, A-20106). .. Following labelling procedures, cells were mounted on glass slides in ProLong Gold antifade (Life Technologies, ).

    Article Title: Apolipoprotein A-I gene transfer exerts immunomodulatory effects and reduces vascular inflammation and fibrosis in ob/ob mice
    Article Snippet: (Splenocyte) mononuclear cell, CD4-, and CD8-T cell proliferation Splenocytes were labeled with 10 μM of succinimidyl ester of carboxyfluorescein diacetate (CFSE Cell Tract™; Invitrogen, Carlbard, CA, USA) to be able to measure cell proliferation, indicative for MNC activation, as described previously [ ]. .. Therefore, splenocytes were cultured in RPMI1640 medium (Invitrogen, Heidelberg, Germany), supplemented with 10 % FBS and 1 % penicillin/streptomycin for 72 h. Then, cells were stained with monoclonal anti-CD4 or anti-CD8 antibodies (BD Biosciences, Franklin Lakes, NJ, USA), followed by flow cytometry on a MACSQuant Analyzer (Miltenyi Biotec, Bergisch Gladbach, Germany) and analysis with FlowJo 8.7. software (Tree Star) for the calculation of the division index, i.e. the average number of cell divisions that the responding cells undergo (i.e., ignores peak 0).

    Purification:

    Article Title: Cancer mutations of the tumor suppressor SPOP disrupt the formation of active, phase-separated compartments
    Article Snippet: GST-SPOPMATH was expressed in BL21 RIPL cells in LB medium, and purified as previously described by Glutathione-sepharose, TEV cleavage during dialysis, ion-exchange by SP column, and SEC ( ). .. Small fractions of each construct were labeled with 4X Oregon Green 488 Carboxylic Acid, Succinimidyl Ester, 5 isomer (Cat #O6147 Thermo Fisher) at 4 ºC overnight.

    Article Title: Cholangiocarcinoma therapy with nanoparticles that combine downregulation of MicroRNA-210 with inhibition of cancer cell invasiveness
    Article Snippet: Succinimidyl ester of Alexa Fluor® 647 carboxylic acid was from Life Technologies (Eugene, OR). .. AlexaFluor 647-labeled PCX polymers (AF647-PCX) were produced according to the manufacturer's instructions and purified by dialysis to remove unreacted dye.

    Article Title: Sulfiredoxin redox-sensitive interaction with S100A4 and non-muscle myosin IIA regulates cancer cell motility
    Article Snippet: .. Purified Srx (C99S) protein was incubated with 5x excess of succinimidyl ester of Alexa® 546 Fluor carboxylic acid (Invitrogen) and S100A4 (S100A4-SSG) purified protein was incubated with 5x excess of succinimidyl ester of QSY®35 acetic acid (Invitrogen) at room temperature, overnight, under constant agitation to label primary amines according to manufacturer’s recommendations. .. The reaction mixture was processed using a BioSpin 6 (BioRad) size-exclusion spin column to remove an excess of unreacted dyes and exchange reaction buffers to 20mM PB (pH 7.4).

    Software:

    Article Title: Bi-Directional Exchange of Membrane Components Occurs during Co-Culture of Mesenchymal Stem Cells and Nucleus Pulposus Cells
    Article Snippet: Assessment of cell fusion Cellular fusion was ascertained by 5,6 carboxyfluorescein diactetate, succinimidyl ester (CFDA; Invitrogen) and SNARF-1 carboxylic acid, acetate, succinimidyl ester (SNARF-1; (Invitrogen). .. To further characterize cellular fusion, CFDA and SNARF positive cells were sorted (BD Biosciences FACS Aria high speed cell sorter with Diva 5 software) as previously described .

    Article Title: Apolipoprotein A-I gene transfer exerts immunomodulatory effects and reduces vascular inflammation and fibrosis in ob/ob mice
    Article Snippet: (Splenocyte) mononuclear cell, CD4-, and CD8-T cell proliferation Splenocytes were labeled with 10 μM of succinimidyl ester of carboxyfluorescein diacetate (CFSE Cell Tract™; Invitrogen, Carlbard, CA, USA) to be able to measure cell proliferation, indicative for MNC activation, as described previously [ ]. .. Therefore, splenocytes were cultured in RPMI1640 medium (Invitrogen, Heidelberg, Germany), supplemented with 10 % FBS and 1 % penicillin/streptomycin for 72 h. Then, cells were stained with monoclonal anti-CD4 or anti-CD8 antibodies (BD Biosciences, Franklin Lakes, NJ, USA), followed by flow cytometry on a MACSQuant Analyzer (Miltenyi Biotec, Bergisch Gladbach, Germany) and analysis with FlowJo 8.7. software (Tree Star) for the calculation of the division index, i.e. the average number of cell divisions that the responding cells undergo (i.e., ignores peak 0).

    In Vitro:

    Article Title: Sulfiredoxin redox-sensitive interaction with S100A4 and non-muscle myosin IIA regulates cancer cell motility
    Article Snippet: Purified Srx (C99S) protein was incubated with 5x excess of succinimidyl ester of Alexa® 546 Fluor carboxylic acid (Invitrogen) and S100A4 (S100A4-SSG) purified protein was incubated with 5x excess of succinimidyl ester of QSY®35 acetic acid (Invitrogen) at room temperature, overnight, under constant agitation to label primary amines according to manufacturer’s recommendations. .. In vitro S-glutathionylation of S100A4 (25μg in 1.0 ml of 10mM PB, pH=7.4) was performed with 50μM PABA/NO and 1mM GSH (room temperature, 1hr, under constant stirring) using procedures previously described ( ).

    Column Chromatography:

    Article Title: The Mammalian Septin MSF Localizes with Microtubules and Is Required for Completion of Cytokinesis
    Article Snippet: Approximately 60 mg of either anti-MSF or anti-Nedd5 IgG was incubated with 6-(tetramethylrhodamine-5-(and-6)-carboxamido)hexanoic acid, succinimidyl ester, or 6-(fluorescein-5-(and-6)-carboxamido)hexanoic acid, succinimidyl ester (Molecular Probes), respectively, in a 1:10 M ratio (protein:dye). .. Labeling was performed according to the manufacturer's directions with the unlabeled conjugate removed from the reaction by column chromatography over a PD-10 column (Amersham).

    Protein Binding:

    Article Title: The focal adhesion kinase inhibitor PF-562,271 impairs primary CD4+ T cell activation
    Article Snippet: Lymphoprep, RPMI-1640, Fetal Bovine Serum, β-mercaptoethanol, Phosphate Buffered Saline, CellQuantiBlue, Triton X-100, PMSF, leupeptin, aprotinin, BD Optilux 384-well tissue culture treated plates, and Greiner 96-well high protein binding capacity plates were obtained from Thermo Fisher Scientific (Waltham, MA). .. Calcein acetoxymethyl ester, carboxyfluorescein diacetate, succinimidyl ester, CD3/CD28-coated Dynabeads, and Alexa-Fluor 680 goat-anti-mouse IgG were obtained from Life Technologies (Carlsbad, CA).

    Nuclear Magnetic Resonance:

    Article Title: Cancer mutations of the tumor suppressor SPOP disrupt the formation of active, phase-separated compartments
    Article Snippet: Small fractions of each construct were labeled with 4X Oregon Green 488 Carboxylic Acid, Succinimidyl Ester, 5 isomer (Cat #O6147 Thermo Fisher) at 4 ºC overnight. .. Protein expressed for NMR experiments were isotopically labeled with 15 N, 13 C by growing cells in M9 minimal media supplemented with 13 C glucose and 15 N ammonium chloride (Cambridge Isotopes). at 37°C until an OD600 = 0.8.

    Produced:

    Article Title: Cholangiocarcinoma therapy with nanoparticles that combine downregulation of MicroRNA-210 with inhibition of cancer cell invasiveness
    Article Snippet: Succinimidyl ester of Alexa Fluor® 647 carboxylic acid was from Life Technologies (Eugene, OR). .. AlexaFluor 647-labeled PCX polymers (AF647-PCX) were produced according to the manufacturer's instructions and purified by dialysis to remove unreacted dye.

    Activation Assay:

    Article Title: Apolipoprotein A-I gene transfer exerts immunomodulatory effects and reduces vascular inflammation and fibrosis in ob/ob mice
    Article Snippet: .. (Splenocyte) mononuclear cell, CD4-, and CD8-T cell proliferation Splenocytes were labeled with 10 μM of succinimidyl ester of carboxyfluorescein diacetate (CFSE Cell Tract™; Invitrogen, Carlbard, CA, USA) to be able to measure cell proliferation, indicative for MNC activation, as described previously [ ]. .. Therefore, splenocytes were cultured in RPMI1640 medium (Invitrogen, Heidelberg, Germany), supplemented with 10 % FBS and 1 % penicillin/streptomycin for 72 h. Then, cells were stained with monoclonal anti-CD4 or anti-CD8 antibodies (BD Biosciences, Franklin Lakes, NJ, USA), followed by flow cytometry on a MACSQuant Analyzer (Miltenyi Biotec, Bergisch Gladbach, Germany) and analysis with FlowJo 8.7. software (Tree Star) for the calculation of the division index, i.e. the average number of cell divisions that the responding cells undergo (i.e., ignores peak 0).

    Construct:

    Article Title: Cancer mutations of the tumor suppressor SPOP disrupt the formation of active, phase-separated compartments
    Article Snippet: .. Small fractions of each construct were labeled with 4X Oregon Green 488 Carboxylic Acid, Succinimidyl Ester, 5 isomer (Cat #O6147 Thermo Fisher) at 4 ºC overnight. .. Un-reacted label was removed from the proteins by PD-10 column (Thermo Fisher 45–000-67).

    Live Cell Imaging:

    Article Title: Multiple roles of filopodial dynamics in particle capture and phagocytosis and phenotypes of Cdc42 and Myo10 deletion
    Article Snippet: The suspension of BioParticles in live cell imaging medium was sonicated to disperse the particles, and 100 μl was pipetted into a μ-slide I chamber before starting time-lapse imaging via spinning disk confocal microscopy. .. The following BioParticles were used: Alexa Fluor 594-conjugated E. coli (K-12 strain) particles (E23370; Thermo Fisher Scientific), Alexa Fluor 594-conjugated zymosan (polysaccharide prepared from the cell wall of S. cerevisiae (bakers' yeast), catalogue number ; Thermo Fisher Scientific) and pHrodo-conjugated zymosan, prepared using unlabeled zymosan (Z2849; Thermo Fisher Scientific) and the succinimidyl ester of pHrodo red ( ; Thermo Fisher Scientific), which has absorbance and emission maxima of 560 and 585 nm, respectively.

    Marker:

    Article Title: Hemagglutinin-based polyanhydride nanovaccines against H5N1 influenza elicit protective virus neutralizing titers and cell-mediated immunity
    Article Snippet: Single cell populations were labeled with 2.5 μM 5-(and-6)- carboxyfluorescein diacetate, succinimidyl ester (5[6]-CFDA, SE; CFSE; Thermo Fisher Scientific, Waltham, MA, USA). .. Single cell populations were labeled with 2.5 μM 5-(and-6)- carboxyfluorescein diacetate, succinimidyl ester (5[6]-CFDA, SE; CFSE; Thermo Fisher Scientific, Waltham, MA, USA).

    FACS:

    Article Title: Bi-Directional Exchange of Membrane Components Occurs during Co-Culture of Mesenchymal Stem Cells and Nucleus Pulposus Cells
    Article Snippet: Assessment of cell fusion Cellular fusion was ascertained by 5,6 carboxyfluorescein diactetate, succinimidyl ester (CFDA; Invitrogen) and SNARF-1 carboxylic acid, acetate, succinimidyl ester (SNARF-1; (Invitrogen). .. To further characterize cellular fusion, CFDA and SNARF positive cells were sorted (BD Biosciences FACS Aria high speed cell sorter with Diva 5 software) as previously described .

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