Structured Review

Becton Dickinson ethylenediaminetetraacetic acid
Ethylenediaminetetraacetic Acid, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 97/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ethylenediaminetetraacetic acid/product/Becton Dickinson
Average 97 stars, based on 25 article reviews
Price from $9.99 to $1999.99
ethylenediaminetetraacetic acid - by Bioz Stars, 2020-04
97/100 stars

Images

Related Articles

Centrifugation:

Article Title: An apoA-I mimetic peptide increases LCAT activity in mice through increasing HDL concentration
Article Snippet: .. Preparation of HDL associated LCAT enzyme Fresh human or mouse blood was drawn into tubes containing potassium EDTA (Becton Dickinson, Franklin lakes, NJ), and plasma was separated immediately by low-speed centrifugation at 2500 rpm (1430g ) for 30 min at 4ºC. .. Size exclusion chromatography was performed at ambient temperature using a Superose-6 10/300 column on a Bio-Rad FPLC system (Hercules, CA).

Cytometry:

Article Title: Hemagglutinin-Dependent Tropism of H5N1 Avian Influenza Virus for Human Endothelial Cells ▿
Article Snippet: Paragraph title: Flow cytometry. ... For E/P-selectin staining, cells were harvested with PBS-A (Invitrogen)-0.94 mM EDTA, washed with Cell Wash (Becton Dickinson), and incubated with CD62E/P antibody (Serotec) for 20 min at 4°C.

Article Title: Hemolytic uremic syndrome-associated Shiga toxins promote endothelial-cell secretion and impair ADAMTS13 cleavage of unusually large von Willebrand factor multimers
Article Snippet: .. HUVECs and GMVECs were detached with 5 mM EDTA (ethylenediaminetetraacetic acid), treated with 0.2% paraformaldehyde at 4°C and 0.2% Tween 20 at 37°C, blocked with 10% human serum/PBS, incubated with either IgM mouse anti-Gb3 (CD77)-fluorescein isothiocyanate (FITC) or irrelevant mouse IgM-FITC (as control), and analyzed by flow cytometry (Becton Dickinson FACScan, San Jose, CA). shows that about 30% to 60% of HUVECs and GMVECs were positive for Gb3 (CD77). .. The flow cytometry data are also shown in .

Article Title: Accumulation of Tissue Factor into Developing Thrombi In Vivo Is Dependent upon Microparticle P-Selectin Glycoprotein Ligand 1 and Platelet P-Selectin
Article Snippet: Beads were incubated with PE-conjugated anti–PSGL-1 antibody at 24°C for 40 min, and then examined for fluorescence on the FACSCalibur® flow cytometer. .. For analysis of PSGL-1 function on microparticles, P-selectin IgG chimera–coated beads were prepared as the antibody-coated beads, which were incubated with calcein-labeled microparticles that had been pretreated with either 2 mM EDTA, 10 μg/ml anti–PSGL-1 monoclonal antibody (BD Biosciences), or 10 μg/ml of its isotype control.

Article Title: Staphylococcus aureus Produces Membrane-Derived Vesicles That Induce Host Cell Death
Article Snippet: .. To determine the cytotoxic effect of lysed MVs, the purified MVs were lysed with 0.1M EDTA at 37°C for 1 h and then treated to HEp-2 cells for 24 h. The samples were immediately analyzed by the flow cytometry and CellQuest Pro software (BD Biosciences). ..

Article Title: Novel ecto-tagged integrins reveal their trafficking in live cells
Article Snippet: .. Mammalian cells were suspended with trypsin/EDTA, incubated with FN9-10 in the presence or absence of 10 mM EDTA or 0.5 mM MnCl2 , fixed with 4% paraformaldehyde, incubated with APC-Strepatavidin and analyzed by flow cytometry on an LSRII instrument (BD Biosciences). .. The activation index of cells was calculated as AI = (F −F o )/F integrin , where F is the geometric mean fluorescence intensity (MFI) of FN9-10 binding, F o is the MFI of FN9-10 binding in presence of inhibitor, and F integrin is the normalized MFI of anti-β1 or anti-α5 integrin antibody binding to cells.

Electrophoresis:

Article Title: Caspase 3-Mediated Inactivation of Rac GTPases Promotes Drug-Induced Apoptosis in Human Lymphoma Cells
Article Snippet: Equal amounts of protein (20 μg per lane) were separated by electrophoresis with a 4 to 12% NuPAGE Bis-Tris gel (Invitrogen), and transferred to nitrocellulose membranes (Millipore) for immunoblotting. .. For in vitro cleavage assays, recombinant His-Rac1 protein (100 ng/μl) was incubated with purified caspase 3 or 8 at a concentration of 5 ng/μl at 37°C in caspase reaction buffer (50 mM HEPES [pH 7.5], 100 mM NaCl, 1 mM EDTA, 10% [wt/vol] sucrose, 0.1% CHAPS, 10 mM dithiothreitol) (BD Biosciences).

Incubation:

Article Title: Caspase 3-Mediated Inactivation of Rac GTPases Promotes Drug-Induced Apoptosis in Human Lymphoma Cells
Article Snippet: .. For in vitro cleavage assays, recombinant His-Rac1 protein (100 ng/μl) was incubated with purified caspase 3 or 8 at a concentration of 5 ng/μl at 37°C in caspase reaction buffer (50 mM HEPES [pH 7.5], 100 mM NaCl, 1 mM EDTA, 10% [wt/vol] sucrose, 0.1% CHAPS, 10 mM dithiothreitol) (BD Biosciences). ..

Article Title: Hemagglutinin-Dependent Tropism of H5N1 Avian Influenza Virus for Human Endothelial Cells ▿
Article Snippet: .. For E/P-selectin staining, cells were harvested with PBS-A (Invitrogen)-0.94 mM EDTA, washed with Cell Wash (Becton Dickinson), and incubated with CD62E/P antibody (Serotec) for 20 min at 4°C. .. Cells were washed and then incubated with fluorescein isothiocyanate-conjugated goat-anti mouse immunoglobulin G1 (IgG1) antibody (Southern Biotech) for 15 min at 4°C before a final wash. For NP detection, cells prepared as described above were fixed and permeabilized with Fix & Perm cell permeabilization kit (An der Grub) according to the manufacturer's instructions and labeled using anti-NP clone HB65 (ATCC).

Article Title: Hemolytic uremic syndrome-associated Shiga toxins promote endothelial-cell secretion and impair ADAMTS13 cleavage of unusually large von Willebrand factor multimers
Article Snippet: .. HUVECs and GMVECs were detached with 5 mM EDTA (ethylenediaminetetraacetic acid), treated with 0.2% paraformaldehyde at 4°C and 0.2% Tween 20 at 37°C, blocked with 10% human serum/PBS, incubated with either IgM mouse anti-Gb3 (CD77)-fluorescein isothiocyanate (FITC) or irrelevant mouse IgM-FITC (as control), and analyzed by flow cytometry (Becton Dickinson FACScan, San Jose, CA). shows that about 30% to 60% of HUVECs and GMVECs were positive for Gb3 (CD77). .. The flow cytometry data are also shown in .

Article Title: Accumulation of Tissue Factor into Developing Thrombi In Vivo Is Dependent upon Microparticle P-Selectin Glycoprotein Ligand 1 and Platelet P-Selectin
Article Snippet: .. For analysis of PSGL-1 function on microparticles, P-selectin IgG chimera–coated beads were prepared as the antibody-coated beads, which were incubated with calcein-labeled microparticles that had been pretreated with either 2 mM EDTA, 10 μg/ml anti–PSGL-1 monoclonal antibody (BD Biosciences), or 10 μg/ml of its isotype control. .. Results We have previously described the use of real-time in vivo fluorescence and brightfield microscopy to capture digital videos of thrombotic events in arterioles in the mouse cremaster microcirculation ( ).

Article Title: Novel ecto-tagged integrins reveal their trafficking in live cells
Article Snippet: .. Mammalian cells were suspended with trypsin/EDTA, incubated with FN9-10 in the presence or absence of 10 mM EDTA or 0.5 mM MnCl2 , fixed with 4% paraformaldehyde, incubated with APC-Strepatavidin and analyzed by flow cytometry on an LSRII instrument (BD Biosciences). .. The activation index of cells was calculated as AI = (F −F o )/F integrin , where F is the geometric mean fluorescence intensity (MFI) of FN9-10 binding, F o is the MFI of FN9-10 binding in presence of inhibitor, and F integrin is the normalized MFI of anti-β1 or anti-α5 integrin antibody binding to cells.

Activity Assay:

Article Title: Evaluation of Anticoagulants for Serologic Assays of Cholera Vaccination
Article Snippet: To examine the direct effect of heparin on vibriocidal activity in serum, 10 human serum samples with low, medium, and high vibriocidal antibody titers were chosen from subjects who had received bivalent, killed whole-cell, oral cholera vaccine (Shanchol) in clinical trials in Kolkata, India. .. Blood (3 ml) was collected twice at day 0 and day 28 using tubes containing EDTA, ACD, or heparin or without anticoagulant (BD Vacutainer; Becton, Dickinson, NJ, USA).

Article Title: Accumulation of Tissue Factor into Developing Thrombi In Vivo Is Dependent upon Microparticle P-Selectin Glycoprotein Ligand 1 and Platelet P-Selectin
Article Snippet: The values were converted to arbitrary units of procoagulant activity by comparison with a standard curve obtained using a rabbit brain thromboplastin standard. .. For analysis of PSGL-1 function on microparticles, P-selectin IgG chimera–coated beads were prepared as the antibody-coated beads, which were incubated with calcein-labeled microparticles that had been pretreated with either 2 mM EDTA, 10 μg/ml anti–PSGL-1 monoclonal antibody (BD Biosciences), or 10 μg/ml of its isotype control.

Expressing:

Article Title: Olive oil bioactives protect pigs against experimentally-induced chronic inflammation independently of alterations in gut microbiota
Article Snippet: Collection of plasma and intestinal samples Three h after the final LPS challenge, blood samples were collected via jugular venipuncture in tubes containing EDTA and aprotinin (BD Vacutainer® ), held in ice-cold water for 30 min, centrifuged at 2,000×g for 10 min, and stored frozen at -80°C until analyses. .. The mucosa was scraped, placed in RNAlater (Applied Biosystems via Thermo Fisher Scientific, Darmstadt, Germany) and then stored at -80°C until gene expression analysis.

Article Title: Hemolytic uremic syndrome-associated Shiga toxins promote endothelial-cell secretion and impair ADAMTS13 cleavage of unusually large von Willebrand factor multimers
Article Snippet: HUVECs and GMVECs were detached with 5 mM EDTA (ethylenediaminetetraacetic acid), treated with 0.2% paraformaldehyde at 4°C and 0.2% Tween 20 at 37°C, blocked with 10% human serum/PBS, incubated with either IgM mouse anti-Gb3 (CD77)-fluorescein isothiocyanate (FITC) or irrelevant mouse IgM-FITC (as control), and analyzed by flow cytometry (Becton Dickinson FACScan, San Jose, CA). shows that about 30% to 60% of HUVECs and GMVECs were positive for Gb3 (CD77). .. It is not possible to make direct quantitative comparisons between the expression of Gb3 on our cultured human endothelial cells in vitro and the expression of Gb3 on human endothelial cells in situ.

Modification:

Article Title: Novel ecto-tagged integrins reveal their trafficking in live cells
Article Snippet: The activation state of endogenous α5β1 integrins was assessed using a modified version of a previously described flow cytometric assay . .. Mammalian cells were suspended with trypsin/EDTA, incubated with FN9-10 in the presence or absence of 10 mM EDTA or 0.5 mM MnCl2 , fixed with 4% paraformaldehyde, incubated with APC-Strepatavidin and analyzed by flow cytometry on an LSRII instrument (BD Biosciences).

Western Blot:

Article Title: Caspase 3-Mediated Inactivation of Rac GTPases Promotes Drug-Induced Apoptosis in Human Lymphoma Cells
Article Snippet: When necessary, the membrane was stripped by Restore Western blot stripping buffer (Pierce) and reprobed with appropriate antibodies. .. For in vitro cleavage assays, recombinant His-Rac1 protein (100 ng/μl) was incubated with purified caspase 3 or 8 at a concentration of 5 ng/μl at 37°C in caspase reaction buffer (50 mM HEPES [pH 7.5], 100 mM NaCl, 1 mM EDTA, 10% [wt/vol] sucrose, 0.1% CHAPS, 10 mM dithiothreitol) (BD Biosciences).

Flow Cytometry:

Article Title: Hemagglutinin-Dependent Tropism of H5N1 Avian Influenza Virus for Human Endothelial Cells ▿
Article Snippet: Paragraph title: Flow cytometry. ... For E/P-selectin staining, cells were harvested with PBS-A (Invitrogen)-0.94 mM EDTA, washed with Cell Wash (Becton Dickinson), and incubated with CD62E/P antibody (Serotec) for 20 min at 4°C.

Article Title: Hemolytic uremic syndrome-associated Shiga toxins promote endothelial-cell secretion and impair ADAMTS13 cleavage of unusually large von Willebrand factor multimers
Article Snippet: .. HUVECs and GMVECs were detached with 5 mM EDTA (ethylenediaminetetraacetic acid), treated with 0.2% paraformaldehyde at 4°C and 0.2% Tween 20 at 37°C, blocked with 10% human serum/PBS, incubated with either IgM mouse anti-Gb3 (CD77)-fluorescein isothiocyanate (FITC) or irrelevant mouse IgM-FITC (as control), and analyzed by flow cytometry (Becton Dickinson FACScan, San Jose, CA). shows that about 30% to 60% of HUVECs and GMVECs were positive for Gb3 (CD77). .. The flow cytometry data are also shown in .

Article Title: An apoA-I mimetic peptide increases LCAT activity in mice through increasing HDL concentration
Article Snippet: Preparation of HDL associated LCAT enzyme Fresh human or mouse blood was drawn into tubes containing potassium EDTA (Becton Dickinson, Franklin lakes, NJ), and plasma was separated immediately by low-speed centrifugation at 2500 rpm (1430g ) for 30 min at 4ºC. .. A 200 µl aliquot of plasma was injected per run, and eluted in PBS with 1 mM EDTA at a flow rate 0.2 ml/min.

Article Title: Accumulation of Tissue Factor into Developing Thrombi In Vivo Is Dependent upon Microparticle P-Selectin Glycoprotein Ligand 1 and Platelet P-Selectin
Article Snippet: Beads were incubated with PE-conjugated anti–PSGL-1 antibody at 24°C for 40 min, and then examined for fluorescence on the FACSCalibur® flow cytometer. .. For analysis of PSGL-1 function on microparticles, P-selectin IgG chimera–coated beads were prepared as the antibody-coated beads, which were incubated with calcein-labeled microparticles that had been pretreated with either 2 mM EDTA, 10 μg/ml anti–PSGL-1 monoclonal antibody (BD Biosciences), or 10 μg/ml of its isotype control.

Article Title: Staphylococcus aureus Produces Membrane-Derived Vesicles That Induce Host Cell Death
Article Snippet: .. To determine the cytotoxic effect of lysed MVs, the purified MVs were lysed with 0.1M EDTA at 37°C for 1 h and then treated to HEp-2 cells for 24 h. The samples were immediately analyzed by the flow cytometry and CellQuest Pro software (BD Biosciences). ..

Article Title: Novel ecto-tagged integrins reveal their trafficking in live cells
Article Snippet: .. Mammalian cells were suspended with trypsin/EDTA, incubated with FN9-10 in the presence or absence of 10 mM EDTA or 0.5 mM MnCl2 , fixed with 4% paraformaldehyde, incubated with APC-Strepatavidin and analyzed by flow cytometry on an LSRII instrument (BD Biosciences). .. The activation index of cells was calculated as AI = (F −F o )/F integrin , where F is the geometric mean fluorescence intensity (MFI) of FN9-10 binding, F o is the MFI of FN9-10 binding in presence of inhibitor, and F integrin is the normalized MFI of anti-β1 or anti-α5 integrin antibody binding to cells.

Concentration Assay:

Article Title: Caspase 3-Mediated Inactivation of Rac GTPases Promotes Drug-Induced Apoptosis in Human Lymphoma Cells
Article Snippet: .. For in vitro cleavage assays, recombinant His-Rac1 protein (100 ng/μl) was incubated with purified caspase 3 or 8 at a concentration of 5 ng/μl at 37°C in caspase reaction buffer (50 mM HEPES [pH 7.5], 100 mM NaCl, 1 mM EDTA, 10% [wt/vol] sucrose, 0.1% CHAPS, 10 mM dithiothreitol) (BD Biosciences). ..

In Vitro:

Article Title: Caspase 3-Mediated Inactivation of Rac GTPases Promotes Drug-Induced Apoptosis in Human Lymphoma Cells
Article Snippet: .. For in vitro cleavage assays, recombinant His-Rac1 protein (100 ng/μl) was incubated with purified caspase 3 or 8 at a concentration of 5 ng/μl at 37°C in caspase reaction buffer (50 mM HEPES [pH 7.5], 100 mM NaCl, 1 mM EDTA, 10% [wt/vol] sucrose, 0.1% CHAPS, 10 mM dithiothreitol) (BD Biosciences). ..

Article Title: Hemolytic uremic syndrome-associated Shiga toxins promote endothelial-cell secretion and impair ADAMTS13 cleavage of unusually large von Willebrand factor multimers
Article Snippet: HUVECs and GMVECs were detached with 5 mM EDTA (ethylenediaminetetraacetic acid), treated with 0.2% paraformaldehyde at 4°C and 0.2% Tween 20 at 37°C, blocked with 10% human serum/PBS, incubated with either IgM mouse anti-Gb3 (CD77)-fluorescein isothiocyanate (FITC) or irrelevant mouse IgM-FITC (as control), and analyzed by flow cytometry (Becton Dickinson FACScan, San Jose, CA). shows that about 30% to 60% of HUVECs and GMVECs were positive for Gb3 (CD77). .. It is not possible to make direct quantitative comparisons between the expression of Gb3 on our cultured human endothelial cells in vitro and the expression of Gb3 on human endothelial cells in situ.

Cell Culture:

Article Title: Hemolytic uremic syndrome-associated Shiga toxins promote endothelial-cell secretion and impair ADAMTS13 cleavage of unusually large von Willebrand factor multimers
Article Snippet: HUVECs and GMVECs were detached with 5 mM EDTA (ethylenediaminetetraacetic acid), treated with 0.2% paraformaldehyde at 4°C and 0.2% Tween 20 at 37°C, blocked with 10% human serum/PBS, incubated with either IgM mouse anti-Gb3 (CD77)-fluorescein isothiocyanate (FITC) or irrelevant mouse IgM-FITC (as control), and analyzed by flow cytometry (Becton Dickinson FACScan, San Jose, CA). shows that about 30% to 60% of HUVECs and GMVECs were positive for Gb3 (CD77). .. It is not possible to make direct quantitative comparisons between the expression of Gb3 on our cultured human endothelial cells in vitro and the expression of Gb3 on human endothelial cells in situ.

other:

Article Title: Presence of C1-Inhibitor Polymers in a Subset of Patients Suffering from Hereditary Angioedema
Article Snippet: Plasma and genotyping of HAE patients EDTA and CPDA plasma samples were collected from 75 Danish HAE patients representing 31 families and 34 healthy individuals (BD Vacutainer K2 EDTA tubes, Becton Dickinson, Plymouth, UK and Vacuette CPDA tubes, Greiner Bio-One, Wemmel, Belgium, respectively).

Injection:

Article Title: An apoA-I mimetic peptide increases LCAT activity in mice through increasing HDL concentration
Article Snippet: Preparation of HDL associated LCAT enzyme Fresh human or mouse blood was drawn into tubes containing potassium EDTA (Becton Dickinson, Franklin lakes, NJ), and plasma was separated immediately by low-speed centrifugation at 2500 rpm (1430g ) for 30 min at 4ºC. .. A 200 µl aliquot of plasma was injected per run, and eluted in PBS with 1 mM EDTA at a flow rate 0.2 ml/min.

Recombinant:

Article Title: Caspase 3-Mediated Inactivation of Rac GTPases Promotes Drug-Induced Apoptosis in Human Lymphoma Cells
Article Snippet: .. For in vitro cleavage assays, recombinant His-Rac1 protein (100 ng/μl) was incubated with purified caspase 3 or 8 at a concentration of 5 ng/μl at 37°C in caspase reaction buffer (50 mM HEPES [pH 7.5], 100 mM NaCl, 1 mM EDTA, 10% [wt/vol] sucrose, 0.1% CHAPS, 10 mM dithiothreitol) (BD Biosciences). ..

Immunofluorescence:

Article Title: Hemolytic uremic syndrome-associated Shiga toxins promote endothelial-cell secretion and impair ADAMTS13 cleavage of unusually large von Willebrand factor multimers
Article Snippet: The cells were initially from decapsulated glomeruli isolated from normal human kidney cortical tissue and were more than 95% positive for cytoplasmic VWF and E-selectin by immunofluorescence. .. HUVECs and GMVECs were detached with 5 mM EDTA (ethylenediaminetetraacetic acid), treated with 0.2% paraformaldehyde at 4°C and 0.2% Tween 20 at 37°C, blocked with 10% human serum/PBS, incubated with either IgM mouse anti-Gb3 (CD77)-fluorescein isothiocyanate (FITC) or irrelevant mouse IgM-FITC (as control), and analyzed by flow cytometry (Becton Dickinson FACScan, San Jose, CA). shows that about 30% to 60% of HUVECs and GMVECs were positive for Gb3 (CD77).

Fluorescence:

Article Title: Accumulation of Tissue Factor into Developing Thrombi In Vivo Is Dependent upon Microparticle P-Selectin Glycoprotein Ligand 1 and Platelet P-Selectin
Article Snippet: Background fluorescence was determined by preincubation of the beads with a 50-fold excess of the same antibody but lacking a fluorescent label before the addition of PE-conjugated anti–PSGL-1 antibody. .. For analysis of PSGL-1 function on microparticles, P-selectin IgG chimera–coated beads were prepared as the antibody-coated beads, which were incubated with calcein-labeled microparticles that had been pretreated with either 2 mM EDTA, 10 μg/ml anti–PSGL-1 monoclonal antibody (BD Biosciences), or 10 μg/ml of its isotype control.

Article Title: Novel ecto-tagged integrins reveal their trafficking in live cells
Article Snippet: Mammalian cells were suspended with trypsin/EDTA, incubated with FN9-10 in the presence or absence of 10 mM EDTA or 0.5 mM MnCl2 , fixed with 4% paraformaldehyde, incubated with APC-Strepatavidin and analyzed by flow cytometry on an LSRII instrument (BD Biosciences). .. The activation index of cells was calculated as AI = (F −F o )/F integrin , where F is the geometric mean fluorescence intensity (MFI) of FN9-10 binding, F o is the MFI of FN9-10 binding in presence of inhibitor, and F integrin is the normalized MFI of anti-β1 or anti-α5 integrin antibody binding to cells.

Isolation:

Article Title: Hemolytic uremic syndrome-associated Shiga toxins promote endothelial-cell secretion and impair ADAMTS13 cleavage of unusually large von Willebrand factor multimers
Article Snippet: The cells were initially from decapsulated glomeruli isolated from normal human kidney cortical tissue and were more than 95% positive for cytoplasmic VWF and E-selectin by immunofluorescence. .. HUVECs and GMVECs were detached with 5 mM EDTA (ethylenediaminetetraacetic acid), treated with 0.2% paraformaldehyde at 4°C and 0.2% Tween 20 at 37°C, blocked with 10% human serum/PBS, incubated with either IgM mouse anti-Gb3 (CD77)-fluorescein isothiocyanate (FITC) or irrelevant mouse IgM-FITC (as control), and analyzed by flow cytometry (Becton Dickinson FACScan, San Jose, CA). shows that about 30% to 60% of HUVECs and GMVECs were positive for Gb3 (CD77).

Size-exclusion Chromatography:

Article Title: An apoA-I mimetic peptide increases LCAT activity in mice through increasing HDL concentration
Article Snippet: Preparation of HDL associated LCAT enzyme Fresh human or mouse blood was drawn into tubes containing potassium EDTA (Becton Dickinson, Franklin lakes, NJ), and plasma was separated immediately by low-speed centrifugation at 2500 rpm (1430g ) for 30 min at 4ºC. .. Size exclusion chromatography was performed at ambient temperature using a Superose-6 10/300 column on a Bio-Rad FPLC system (Hercules, CA).

Labeling:

Article Title: Hemagglutinin-Dependent Tropism of H5N1 Avian Influenza Virus for Human Endothelial Cells ▿
Article Snippet: For E/P-selectin staining, cells were harvested with PBS-A (Invitrogen)-0.94 mM EDTA, washed with Cell Wash (Becton Dickinson), and incubated with CD62E/P antibody (Serotec) for 20 min at 4°C. .. Cells were washed and then incubated with fluorescein isothiocyanate-conjugated goat-anti mouse immunoglobulin G1 (IgG1) antibody (Southern Biotech) for 15 min at 4°C before a final wash. For NP detection, cells prepared as described above were fixed and permeabilized with Fix & Perm cell permeabilization kit (An der Grub) according to the manufacturer's instructions and labeled using anti-NP clone HB65 (ATCC).

Purification:

Article Title: Caspase 3-Mediated Inactivation of Rac GTPases Promotes Drug-Induced Apoptosis in Human Lymphoma Cells
Article Snippet: .. For in vitro cleavage assays, recombinant His-Rac1 protein (100 ng/μl) was incubated with purified caspase 3 or 8 at a concentration of 5 ng/μl at 37°C in caspase reaction buffer (50 mM HEPES [pH 7.5], 100 mM NaCl, 1 mM EDTA, 10% [wt/vol] sucrose, 0.1% CHAPS, 10 mM dithiothreitol) (BD Biosciences). ..

Article Title: Accumulation of Tissue Factor into Developing Thrombi In Vivo Is Dependent upon Microparticle P-Selectin Glycoprotein Ligand 1 and Platelet P-Selectin
Article Snippet: Paragraph title: Purification and Analysis of Tissue Factor and PSGL-1–bearing Microparticles ... For analysis of PSGL-1 function on microparticles, P-selectin IgG chimera–coated beads were prepared as the antibody-coated beads, which were incubated with calcein-labeled microparticles that had been pretreated with either 2 mM EDTA, 10 μg/ml anti–PSGL-1 monoclonal antibody (BD Biosciences), or 10 μg/ml of its isotype control.

Article Title: Staphylococcus aureus Produces Membrane-Derived Vesicles That Induce Host Cell Death
Article Snippet: .. To determine the cytotoxic effect of lysed MVs, the purified MVs were lysed with 0.1M EDTA at 37°C for 1 h and then treated to HEp-2 cells for 24 h. The samples were immediately analyzed by the flow cytometry and CellQuest Pro software (BD Biosciences). ..

Article Title: Novel ecto-tagged integrins reveal their trafficking in live cells
Article Snippet: Briefly, human FN repeats 9-10 fused to an N-terminal His-tagged Maltose-Binding Protein affinity tag, was purified from E. Coli and biotinylated with N -hydroxysuccinimidobiotin (ThermoFisher Scientific; FN9-10; manuscript in preparation). .. Mammalian cells were suspended with trypsin/EDTA, incubated with FN9-10 in the presence or absence of 10 mM EDTA or 0.5 mM MnCl2 , fixed with 4% paraformaldehyde, incubated with APC-Strepatavidin and analyzed by flow cytometry on an LSRII instrument (BD Biosciences).

Stripping:

Article Title: Caspase 3-Mediated Inactivation of Rac GTPases Promotes Drug-Induced Apoptosis in Human Lymphoma Cells
Article Snippet: When necessary, the membrane was stripped by Restore Western blot stripping buffer (Pierce) and reprobed with appropriate antibodies. .. For in vitro cleavage assays, recombinant His-Rac1 protein (100 ng/μl) was incubated with purified caspase 3 or 8 at a concentration of 5 ng/μl at 37°C in caspase reaction buffer (50 mM HEPES [pH 7.5], 100 mM NaCl, 1 mM EDTA, 10% [wt/vol] sucrose, 0.1% CHAPS, 10 mM dithiothreitol) (BD Biosciences).

Fast Protein Liquid Chromatography:

Article Title: An apoA-I mimetic peptide increases LCAT activity in mice through increasing HDL concentration
Article Snippet: Preparation of HDL associated LCAT enzyme Fresh human or mouse blood was drawn into tubes containing potassium EDTA (Becton Dickinson, Franklin lakes, NJ), and plasma was separated immediately by low-speed centrifugation at 2500 rpm (1430g ) for 30 min at 4ºC. .. Size exclusion chromatography was performed at ambient temperature using a Superose-6 10/300 column on a Bio-Rad FPLC system (Hercules, CA).

Lysis:

Article Title: Caspase 3-Mediated Inactivation of Rac GTPases Promotes Drug-Induced Apoptosis in Human Lymphoma Cells
Article Snippet: Drug-treated cells were washed twice in ice-cold phosphate-buffered saline and lysed in lysis buffer (25 mM Tris-HCl [pH 6.8], 2% SDS, and 10% glycerol). .. For in vitro cleavage assays, recombinant His-Rac1 protein (100 ng/μl) was incubated with purified caspase 3 or 8 at a concentration of 5 ng/μl at 37°C in caspase reaction buffer (50 mM HEPES [pH 7.5], 100 mM NaCl, 1 mM EDTA, 10% [wt/vol] sucrose, 0.1% CHAPS, 10 mM dithiothreitol) (BD Biosciences).

Software:

Article Title: Effect of Sipjeondaebo-Tang on Cancer-Induced Anorexia and Cachexia in CT-26 Tumor-Bearing Mice
Article Snippet: Standards were plotted and concentrations were determined using Milliplex Analyst software version 5.1. .. The blood samples were placed in Vacutainer TM tubes containing EDTA (BD Science, NJ, USA).

Article Title: Accumulation of Tissue Factor into Developing Thrombi In Vivo Is Dependent upon Microparticle P-Selectin Glycoprotein Ligand 1 and Platelet P-Selectin
Article Snippet: Data were analyzed using CellQuest™ software. .. For analysis of PSGL-1 function on microparticles, P-selectin IgG chimera–coated beads were prepared as the antibody-coated beads, which were incubated with calcein-labeled microparticles that had been pretreated with either 2 mM EDTA, 10 μg/ml anti–PSGL-1 monoclonal antibody (BD Biosciences), or 10 μg/ml of its isotype control.

Article Title: Staphylococcus aureus Produces Membrane-Derived Vesicles That Induce Host Cell Death
Article Snippet: .. To determine the cytotoxic effect of lysed MVs, the purified MVs were lysed with 0.1M EDTA at 37°C for 1 h and then treated to HEp-2 cells for 24 h. The samples were immediately analyzed by the flow cytometry and CellQuest Pro software (BD Biosciences). ..

Article Title: Novel ecto-tagged integrins reveal their trafficking in live cells
Article Snippet: Mammalian cells were suspended with trypsin/EDTA, incubated with FN9-10 in the presence or absence of 10 mM EDTA or 0.5 mM MnCl2 , fixed with 4% paraformaldehyde, incubated with APC-Strepatavidin and analyzed by flow cytometry on an LSRII instrument (BD Biosciences). .. Flow cytometry data were analyzed using FlowJo software.

Binding Assay:

Article Title: Staphylococcus aureus Produces Membrane-Derived Vesicles That Induce Host Cell Death
Article Snippet: The cells were stained with FITC-conjugated Annexin V in Annexin V binding buffer for 15 min. PI was added to determine alterations in cell membrane integrity. .. To determine the cytotoxic effect of lysed MVs, the purified MVs were lysed with 0.1M EDTA at 37°C for 1 h and then treated to HEp-2 cells for 24 h. The samples were immediately analyzed by the flow cytometry and CellQuest Pro software (BD Biosciences).

Article Title: Novel ecto-tagged integrins reveal their trafficking in live cells
Article Snippet: Mammalian cells were suspended with trypsin/EDTA, incubated with FN9-10 in the presence or absence of 10 mM EDTA or 0.5 mM MnCl2 , fixed with 4% paraformaldehyde, incubated with APC-Strepatavidin and analyzed by flow cytometry on an LSRII instrument (BD Biosciences). .. The activation index of cells was calculated as AI = (F −F o )/F integrin , where F is the geometric mean fluorescence intensity (MFI) of FN9-10 binding, F o is the MFI of FN9-10 binding in presence of inhibitor, and F integrin is the normalized MFI of anti-β1 or anti-α5 integrin antibody binding to cells.

In Situ:

Article Title: Hemolytic uremic syndrome-associated Shiga toxins promote endothelial-cell secretion and impair ADAMTS13 cleavage of unusually large von Willebrand factor multimers
Article Snippet: HUVECs and GMVECs were detached with 5 mM EDTA (ethylenediaminetetraacetic acid), treated with 0.2% paraformaldehyde at 4°C and 0.2% Tween 20 at 37°C, blocked with 10% human serum/PBS, incubated with either IgM mouse anti-Gb3 (CD77)-fluorescein isothiocyanate (FITC) or irrelevant mouse IgM-FITC (as control), and analyzed by flow cytometry (Becton Dickinson FACScan, San Jose, CA). shows that about 30% to 60% of HUVECs and GMVECs were positive for Gb3 (CD77). .. It is not possible to make direct quantitative comparisons between the expression of Gb3 on our cultured human endothelial cells in vitro and the expression of Gb3 on human endothelial cells in situ.

Sampling:

Article Title: Development and validation of a clinical HPLC method for the quantification of hydroxychloroquine and its metabolites in whole blood
Article Snippet: Blood collection To investigate the effect of anticoagulant on HCQ assay, whole blood samples from 65 SLE patients treated with HCQ were collected in both EDTA and lithium-heparinate tubes (Becton Dickinson, NJ, USA) without gel separator. .. All patients agreed for the sampling analysis in compliance with French regulations.

Activation Assay:

Article Title: Novel ecto-tagged integrins reveal their trafficking in live cells
Article Snippet: Paragraph title: Flow cytometry and analysis of integrin activation ... Mammalian cells were suspended with trypsin/EDTA, incubated with FN9-10 in the presence or absence of 10 mM EDTA or 0.5 mM MnCl2 , fixed with 4% paraformaldehyde, incubated with APC-Strepatavidin and analyzed by flow cytometry on an LSRII instrument (BD Biosciences).

Coagulation:

Article Title: Accumulation of Tissue Factor into Developing Thrombi In Vivo Is Dependent upon Microparticle P-Selectin Glycoprotein Ligand 1 and Platelet P-Selectin
Article Snippet: Tissue factor activity was assayed with a one stage clotting time ( ). .. For analysis of PSGL-1 function on microparticles, P-selectin IgG chimera–coated beads were prepared as the antibody-coated beads, which were incubated with calcein-labeled microparticles that had been pretreated with either 2 mM EDTA, 10 μg/ml anti–PSGL-1 monoclonal antibody (BD Biosciences), or 10 μg/ml of its isotype control.

Staining:

Article Title: Hemagglutinin-Dependent Tropism of H5N1 Avian Influenza Virus for Human Endothelial Cells ▿
Article Snippet: .. For E/P-selectin staining, cells were harvested with PBS-A (Invitrogen)-0.94 mM EDTA, washed with Cell Wash (Becton Dickinson), and incubated with CD62E/P antibody (Serotec) for 20 min at 4°C. .. Cells were washed and then incubated with fluorescein isothiocyanate-conjugated goat-anti mouse immunoglobulin G1 (IgG1) antibody (Southern Biotech) for 15 min at 4°C before a final wash. For NP detection, cells prepared as described above were fixed and permeabilized with Fix & Perm cell permeabilization kit (An der Grub) according to the manufacturer's instructions and labeled using anti-NP clone HB65 (ATCC).

Article Title: Staphylococcus aureus Produces Membrane-Derived Vesicles That Induce Host Cell Death
Article Snippet: The cells were stained with FITC-conjugated Annexin V in Annexin V binding buffer for 15 min. PI was added to determine alterations in cell membrane integrity. .. To determine the cytotoxic effect of lysed MVs, the purified MVs were lysed with 0.1M EDTA at 37°C for 1 h and then treated to HEp-2 cells for 24 h. The samples were immediately analyzed by the flow cytometry and CellQuest Pro software (BD Biosciences).

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96
    Becton Dickinson edta microtube
    Scatter plots and Bland-Altman plots comparing the reference CD4 counts versus Pima CD4. Scatter and Bland-Altman plots for capillary blood directly applied to CD4 cartridges, Pima-D (1A and 1B), capillary blood collected in <t>EDTA</t> <t>microtube,</t> Pima-M (2A and 2B), venous blood,Pima-V (3A and 3B), or venous blood tested with the Pima at the reference laboratory, Pima-Lab (4A and 4B) with reference CD4 assay being BD Multitest reagent and BD Trucount Tubes using a BD FACSCalibur.
    Edta Microtube, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 96/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/edta microtube/product/Becton Dickinson
    Average 96 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    edta microtube - by Bioz Stars, 2020-04
    96/100 stars
      Buy from Supplier

    99
    Becton Dickinson facs buffer
    Potential molecular mediators of CD11b + Gr1 + cells for immune regulatory function. CD11b + Gr1 hi cells were purified from the spleen and enzyme activity or level of transcripts were determined. (A) MPO enzyme activity. (B) RT-PCR analysis of arginase gene. (C) NE enzyme activity. DATA pooled from 2 independent experiments for A~C. Representative oncostain M (D) and lactoferrin (E) expression in splenocytes from septic mice determined by immunohistochemistry after treatment with <t>PBS</t> or TDCA. Data shown are representative of 3 independent experiments with 3 samples per experiment. (F) Representative <t>FACS</t> plots of (G) indicating the surface expression of CD244 on splenic CD11b + Gr1 hi cells. The gating strategies are shown in Figure S15 . (G) MFIs of CD244 expression on CD11b + Gr1 hi cells are plotted. Data pooled from 4 independent experiments with 2~4 samples per experiment. (H) Survival rate of septic mice given LPS i.p. after recieving isotype control antibody + TDCA (•), anti-CD244 antibody + TDCA (°), or isotype control antibody + PBS (■). n = 8 (I) Survival rate of septic mice that received LPS i.p. after being administered isotype control antibody + adoptive transfer of MDSC LT (•, n = 10), anti-CD244 antibody + adoptive transfer of MDSC LT (°, n = 12), or isotype control antibody + PBS (■, n = 10). The number of CD11b + Gr1 hi cells in the bone marrow (J) or spleen (K) after adoptive transfer of MDSC L or MDSC LT . The number of total BM cells or total splenocytes were counted and the number of CD11b + Gr1 hi cells was calculated using the % of cells after FACS analysis as in Figure 2E . (L) RT-PCR analysis of prokineticin 2. n = 4, RU, relative units of band intensity normalized to the intensity of GAPDH. Short horizontal lines among the circles indicate the mean values. * p
    Facs Buffer, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 99/100, based on 3656 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/facs buffer/product/Becton Dickinson
    Average 99 stars, based on 3656 article reviews
    Price from $9.99 to $1999.99
    facs buffer - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    97
    Becton Dickinson edta vacutainer tubes
    Kinetic analysis of circulating lymphocytes following influenza virus infection. Three ferrets each were inoculated i.n. with 10 6 PFU of virus. Blood was collected on days 3 (A), 7 (B), and 14 to 16 (C) p.i. in <t>EDTA</t> <t>Vacutainer</t> tubes and analyzed with
    Edta Vacutainer Tubes, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 97/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/edta vacutainer tubes/product/Becton Dickinson
    Average 97 stars, based on 41 article reviews
    Price from $9.99 to $1999.99
    edta vacutainer tubes - by Bioz Stars, 2020-04
    97/100 stars
      Buy from Supplier

    93
    Becton Dickinson potassium edta
    Preparation of HDL associated <t>LCAT.</t> Size exclusion chromatography was perfo rmed at ambient temperature using a Superose-6 10/300 column on a Bio-Rad FPLC system (Hercules, CA). A 200 µl aliquot of plasma was injected per run, and eluted in PBS with 1 mM <t>EDTA</t> at a flow rate of 0.2 ml/min. Total cholesterol in each fraction was measured using the Cholesterol E Kit. LCAT activity was determined as following: A. 20 µl of each fraction of human plasma were incubated at 37ºC for 5 h; B. 2.5 µl of each fraction of WT mouse plasma were incubated at 37ºC for 2 h; C. 10 µl of each fraction of apoE-null mouse plasma were incubated at 37ºC for 4 h.
    Potassium Edta, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/potassium edta/product/Becton Dickinson
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    potassium edta - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    Image Search Results


    Scatter plots and Bland-Altman plots comparing the reference CD4 counts versus Pima CD4. Scatter and Bland-Altman plots for capillary blood directly applied to CD4 cartridges, Pima-D (1A and 1B), capillary blood collected in EDTA microtube, Pima-M (2A and 2B), venous blood,Pima-V (3A and 3B), or venous blood tested with the Pima at the reference laboratory, Pima-Lab (4A and 4B) with reference CD4 assay being BD Multitest reagent and BD Trucount Tubes using a BD FACSCalibur.

    Journal: PLoS ONE

    Article Title: Evaluation of Specimen Types for Pima CD4 Point-of-Care Testing: Advantages of Fingerstick Blood Collection into an EDTA Microtube

    doi: 10.1371/journal.pone.0202018

    Figure Lengend Snippet: Scatter plots and Bland-Altman plots comparing the reference CD4 counts versus Pima CD4. Scatter and Bland-Altman plots for capillary blood directly applied to CD4 cartridges, Pima-D (1A and 1B), capillary blood collected in EDTA microtube, Pima-M (2A and 2B), venous blood,Pima-V (3A and 3B), or venous blood tested with the Pima at the reference laboratory, Pima-Lab (4A and 4B) with reference CD4 assay being BD Multitest reagent and BD Trucount Tubes using a BD FACSCalibur.

    Article Snippet: Fingerstick blood collected in an EDTA microtube allows for multiple testing and reflex testing without additional blood collection.

    Techniques:

    Potential molecular mediators of CD11b + Gr1 + cells for immune regulatory function. CD11b + Gr1 hi cells were purified from the spleen and enzyme activity or level of transcripts were determined. (A) MPO enzyme activity. (B) RT-PCR analysis of arginase gene. (C) NE enzyme activity. DATA pooled from 2 independent experiments for A~C. Representative oncostain M (D) and lactoferrin (E) expression in splenocytes from septic mice determined by immunohistochemistry after treatment with PBS or TDCA. Data shown are representative of 3 independent experiments with 3 samples per experiment. (F) Representative FACS plots of (G) indicating the surface expression of CD244 on splenic CD11b + Gr1 hi cells. The gating strategies are shown in Figure S15 . (G) MFIs of CD244 expression on CD11b + Gr1 hi cells are plotted. Data pooled from 4 independent experiments with 2~4 samples per experiment. (H) Survival rate of septic mice given LPS i.p. after recieving isotype control antibody + TDCA (•), anti-CD244 antibody + TDCA (°), or isotype control antibody + PBS (■). n = 8 (I) Survival rate of septic mice that received LPS i.p. after being administered isotype control antibody + adoptive transfer of MDSC LT (•, n = 10), anti-CD244 antibody + adoptive transfer of MDSC LT (°, n = 12), or isotype control antibody + PBS (■, n = 10). The number of CD11b + Gr1 hi cells in the bone marrow (J) or spleen (K) after adoptive transfer of MDSC L or MDSC LT . The number of total BM cells or total splenocytes were counted and the number of CD11b + Gr1 hi cells was calculated using the % of cells after FACS analysis as in Figure 2E . (L) RT-PCR analysis of prokineticin 2. n = 4, RU, relative units of band intensity normalized to the intensity of GAPDH. Short horizontal lines among the circles indicate the mean values. * p

    Journal: Frontiers in Immunology

    Article Title: Taurodeoxycholate Increases the Number of Myeloid-Derived Suppressor Cells That Ameliorate Sepsis in Mice

    doi: 10.3389/fimmu.2018.01984

    Figure Lengend Snippet: Potential molecular mediators of CD11b + Gr1 + cells for immune regulatory function. CD11b + Gr1 hi cells were purified from the spleen and enzyme activity or level of transcripts were determined. (A) MPO enzyme activity. (B) RT-PCR analysis of arginase gene. (C) NE enzyme activity. DATA pooled from 2 independent experiments for A~C. Representative oncostain M (D) and lactoferrin (E) expression in splenocytes from septic mice determined by immunohistochemistry after treatment with PBS or TDCA. Data shown are representative of 3 independent experiments with 3 samples per experiment. (F) Representative FACS plots of (G) indicating the surface expression of CD244 on splenic CD11b + Gr1 hi cells. The gating strategies are shown in Figure S15 . (G) MFIs of CD244 expression on CD11b + Gr1 hi cells are plotted. Data pooled from 4 independent experiments with 2~4 samples per experiment. (H) Survival rate of septic mice given LPS i.p. after recieving isotype control antibody + TDCA (•), anti-CD244 antibody + TDCA (°), or isotype control antibody + PBS (■). n = 8 (I) Survival rate of septic mice that received LPS i.p. after being administered isotype control antibody + adoptive transfer of MDSC LT (•, n = 10), anti-CD244 antibody + adoptive transfer of MDSC LT (°, n = 12), or isotype control antibody + PBS (■, n = 10). The number of CD11b + Gr1 hi cells in the bone marrow (J) or spleen (K) after adoptive transfer of MDSC L or MDSC LT . The number of total BM cells or total splenocytes were counted and the number of CD11b + Gr1 hi cells was calculated using the % of cells after FACS analysis as in Figure 2E . (L) RT-PCR analysis of prokineticin 2. n = 4, RU, relative units of band intensity normalized to the intensity of GAPDH. Short horizontal lines among the circles indicate the mean values. * p

    Article Snippet: After depleting red blood cells (RBCs) using ACK lysis buffer (0.15 M NH4 Cl, 0.1 mM EDTA, 1 mM Tris pH 7.4), the splenocytes or bone marrow cells were washed with ice-cold FACS buffer (PBS that contained 1% BSA, 0.1% NaN3 and 1 mM EDTA) and blocked on ice for 30 min with blocking buffer [anti-FcγRIIb antibody (clone 2.4G2, BD) and 10% heat-inactivated mixed serum from mouse, rat and goat].

    Techniques: Purification, Activity Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Mouse Assay, Immunohistochemistry, FACS, Adoptive Transfer Assay

    Phenotype of splenic CD11b + Gr1 hi cells increased by TDCA. (A) The absolute numbers of spleen cells from B6 mice at 48 h post LPS injection (left, Data pooled from 5 independent experiments) or 72 h post CLP (right) by Trypan blue exclusion assay. (B) The absolute number of splenic CD11b + Gr1 hi cells from B6 mice at 48 h post LPS injection (left) or 72 h post CLP setting (right). (C) Representative FACS plots of (B) . (D) Representative FACS plots showing the expression of Ly6g and Ly6c on the subpopulation of CD11b + cells. The percentage denotes the % of Ly6c int Ly6g + cells among CD11b + cells. (E) Representative FACS plots showing the expression of Gr1 and F4/80 on CD11b + cells. (F) Representative FACS plots showing expression of CD31 on CD11b + Gr1 hi cells (solid line) and CD11b + Gr1 int cells (dotted line) from the LPS+PBS group and LPS+TDCA group. Filled histograms denote the isotype control. (G) Expression of CD177 on CD11b + Gr1 hi cells. Filled histograms denote the isotype control. (H) [H 3 ]Thymidine-incorporation of T-cells following CD3 + CD28 ligation was measured after co-culture with CD11b + Gr1 hi cells (E:T = 1:5) purified from spleen of mice injected with PBS + PBS, LPS + PBS, or LPS + TDCA. The data are expressed as the mean count per minute (cpm) ± SEM of triplicate assays pooled from 3 independent experiments. (I) The survival of mice after adoptive transfer of CD11b + Gr1 hi cells (1 × 10 5 cells/mouse) purified from the LPS + TDCA group (■, n = 7, MDSC LT ) or the LPS + PBS group (□, n = 5, MDSC L ). CD11b + Gr1 int cells (▴, n = 3) were purified from mice in the LPS + TDCA group. The thin horizontal lines on the top of the FACS plots indicate the gates used in the FACS analysis, and the thick horizontal lines are the experimental groups treated with LPS or PBS. The gating strategies are shown in Figures S7~S12. Data shown are pooled from 3 independent experiments with 1~3 mice per experiment otherwise denoted. Representative FACS plots of 3 independent experiments. † p

    Journal: Frontiers in Immunology

    Article Title: Taurodeoxycholate Increases the Number of Myeloid-Derived Suppressor Cells That Ameliorate Sepsis in Mice

    doi: 10.3389/fimmu.2018.01984

    Figure Lengend Snippet: Phenotype of splenic CD11b + Gr1 hi cells increased by TDCA. (A) The absolute numbers of spleen cells from B6 mice at 48 h post LPS injection (left, Data pooled from 5 independent experiments) or 72 h post CLP (right) by Trypan blue exclusion assay. (B) The absolute number of splenic CD11b + Gr1 hi cells from B6 mice at 48 h post LPS injection (left) or 72 h post CLP setting (right). (C) Representative FACS plots of (B) . (D) Representative FACS plots showing the expression of Ly6g and Ly6c on the subpopulation of CD11b + cells. The percentage denotes the % of Ly6c int Ly6g + cells among CD11b + cells. (E) Representative FACS plots showing the expression of Gr1 and F4/80 on CD11b + cells. (F) Representative FACS plots showing expression of CD31 on CD11b + Gr1 hi cells (solid line) and CD11b + Gr1 int cells (dotted line) from the LPS+PBS group and LPS+TDCA group. Filled histograms denote the isotype control. (G) Expression of CD177 on CD11b + Gr1 hi cells. Filled histograms denote the isotype control. (H) [H 3 ]Thymidine-incorporation of T-cells following CD3 + CD28 ligation was measured after co-culture with CD11b + Gr1 hi cells (E:T = 1:5) purified from spleen of mice injected with PBS + PBS, LPS + PBS, or LPS + TDCA. The data are expressed as the mean count per minute (cpm) ± SEM of triplicate assays pooled from 3 independent experiments. (I) The survival of mice after adoptive transfer of CD11b + Gr1 hi cells (1 × 10 5 cells/mouse) purified from the LPS + TDCA group (■, n = 7, MDSC LT ) or the LPS + PBS group (□, n = 5, MDSC L ). CD11b + Gr1 int cells (▴, n = 3) were purified from mice in the LPS + TDCA group. The thin horizontal lines on the top of the FACS plots indicate the gates used in the FACS analysis, and the thick horizontal lines are the experimental groups treated with LPS or PBS. The gating strategies are shown in Figures S7~S12. Data shown are pooled from 3 independent experiments with 1~3 mice per experiment otherwise denoted. Representative FACS plots of 3 independent experiments. † p

    Article Snippet: After depleting red blood cells (RBCs) using ACK lysis buffer (0.15 M NH4 Cl, 0.1 mM EDTA, 1 mM Tris pH 7.4), the splenocytes or bone marrow cells were washed with ice-cold FACS buffer (PBS that contained 1% BSA, 0.1% NaN3 and 1 mM EDTA) and blocked on ice for 30 min with blocking buffer [anti-FcγRIIb antibody (clone 2.4G2, BD) and 10% heat-inactivated mixed serum from mouse, rat and goat].

    Techniques: Mouse Assay, Injection, Trypan Blue Exclusion Assay, FACS, Expressing, Ligation, Co-Culture Assay, Purification, Adoptive Transfer Assay

    Comparison of gene expression profiles of MDSC L and MDSC LT . (A) CD11b + Gr1 hi cells were FACS-purified from the LPS + PBS group (8 independent sorting, 4~9 mice samples were pooled for each sorting) and the LPS + TDCA group (5 independent sorting, 4~6 mice samples were pooled for each sorting). cDNAs from each mouse were pooled and gene expression was profiled in duplicate. Eight hundred eighteen genes were filtered ( > 8-fold change, p

    Journal: Frontiers in Immunology

    Article Title: Taurodeoxycholate Increases the Number of Myeloid-Derived Suppressor Cells That Ameliorate Sepsis in Mice

    doi: 10.3389/fimmu.2018.01984

    Figure Lengend Snippet: Comparison of gene expression profiles of MDSC L and MDSC LT . (A) CD11b + Gr1 hi cells were FACS-purified from the LPS + PBS group (8 independent sorting, 4~9 mice samples were pooled for each sorting) and the LPS + TDCA group (5 independent sorting, 4~6 mice samples were pooled for each sorting). cDNAs from each mouse were pooled and gene expression was profiled in duplicate. Eight hundred eighteen genes were filtered ( > 8-fold change, p

    Article Snippet: After depleting red blood cells (RBCs) using ACK lysis buffer (0.15 M NH4 Cl, 0.1 mM EDTA, 1 mM Tris pH 7.4), the splenocytes or bone marrow cells were washed with ice-cold FACS buffer (PBS that contained 1% BSA, 0.1% NaN3 and 1 mM EDTA) and blocked on ice for 30 min with blocking buffer [anti-FcγRIIb antibody (clone 2.4G2, BD) and 10% heat-inactivated mixed serum from mouse, rat and goat].

    Techniques: Expressing, FACS, Purification, Mouse Assay

    Kinetic analysis of circulating lymphocytes following influenza virus infection. Three ferrets each were inoculated i.n. with 10 6 PFU of virus. Blood was collected on days 3 (A), 7 (B), and 14 to 16 (C) p.i. in EDTA Vacutainer tubes and analyzed with

    Journal: Journal of Virology

    Article Title: Pathogenesis and Transmission of Triple-Reassortant Swine H1N1 Influenza Viruses Isolated before the 2009 H1N1 Pandemic ▿

    doi: 10.1128/JVI.02231-10

    Figure Lengend Snippet: Kinetic analysis of circulating lymphocytes following influenza virus infection. Three ferrets each were inoculated i.n. with 10 6 PFU of virus. Blood was collected on days 3 (A), 7 (B), and 14 to 16 (C) p.i. in EDTA Vacutainer tubes and analyzed with

    Article Snippet: Complete blood counts were quantified from blood collected in EDTA Vacutainer tubes (BD, Franklin Lakes, NJ) using a Hemavet HV950FS instrument per the manufacturer's instructions (Drew Scientific, Inc., Oxford, CT).

    Techniques: Infection

    Preparation of HDL associated LCAT. Size exclusion chromatography was perfo rmed at ambient temperature using a Superose-6 10/300 column on a Bio-Rad FPLC system (Hercules, CA). A 200 µl aliquot of plasma was injected per run, and eluted in PBS with 1 mM EDTA at a flow rate of 0.2 ml/min. Total cholesterol in each fraction was measured using the Cholesterol E Kit. LCAT activity was determined as following: A. 20 µl of each fraction of human plasma were incubated at 37ºC for 5 h; B. 2.5 µl of each fraction of WT mouse plasma were incubated at 37ºC for 2 h; C. 10 µl of each fraction of apoE-null mouse plasma were incubated at 37ºC for 4 h.

    Journal: International Journal of Biological Sciences

    Article Title: An apoA-I mimetic peptide increases LCAT activity in mice through increasing HDL concentration

    doi:

    Figure Lengend Snippet: Preparation of HDL associated LCAT. Size exclusion chromatography was perfo rmed at ambient temperature using a Superose-6 10/300 column on a Bio-Rad FPLC system (Hercules, CA). A 200 µl aliquot of plasma was injected per run, and eluted in PBS with 1 mM EDTA at a flow rate of 0.2 ml/min. Total cholesterol in each fraction was measured using the Cholesterol E Kit. LCAT activity was determined as following: A. 20 µl of each fraction of human plasma were incubated at 37ºC for 5 h; B. 2.5 µl of each fraction of WT mouse plasma were incubated at 37ºC for 2 h; C. 10 µl of each fraction of apoE-null mouse plasma were incubated at 37ºC for 4 h.

    Article Snippet: Preparation of HDL associated LCAT enzyme Fresh human or mouse blood was drawn into tubes containing potassium EDTA (Becton Dickinson, Franklin lakes, NJ), and plasma was separated immediately by low-speed centrifugation at 2500 rpm (1430g ) for 30 min at 4ºC.

    Techniques: Size-exclusion Chromatography, Fast Protein Liquid Chromatography, Injection, Flow Cytometry, Activity Assay, Incubation