Mediastinal lymph nodes were collected and perfused with dissociation enzyme mix (Miltenyi Biotech, Auburn, CA), finely chopped and incubated for 20 minutes at 37 °C. After enzymatic digestion and red blood cell lysis, samples were incubated with an antibody cocktail consisting of: CD4 FITC, CD3 Alexa Fluor 700, CD25 APC/Cy7 (eBioscience, San Diego, CA) and CD8a BUV 737 (BD Biosciences, San Jose, CA). Cells were then permeabilized and incubated with another antibody cocktail consisting of intracellular markers Foxp3 PE, RORγt PerCP/Cy5.5, T-bet Alexa Fluor 647 (BD Biosciences, San Jose, CA) and GATA-3 PE/Cy7 (R&D Systems, Minneapolis, MN). Cells were analyzed by FACS to determine T-cell subsets in the lymph nodes.
Cd8a buv 737
Manufactured by BD
The CD8a BUV 737 is a fluorochrome-conjugated antibody that binds to the CD8a protein, which is expressed on the surface of certain T cells. This product can be used in flow cytometry applications to identify and quantify CD8a-positive cell populations.
Automatically generated - may contain errors
Lab products found in correlation
Dissociation enzyme mix, by Miltenyi Biotec (1 mentions)
Cd25 apccy7, by Thermo Fisher Scientific (1 mentions)
Rorγt percp cy5, by BD (1 mentions)
Cd3 alexa fluor 700, by Thermo Fisher Scientific (1 mentions)
Cd4 fitc, by Thermo Fisher Scientific (1 mentions)
Cd11b bv421, by BioLegend (1 mentions)
Gr1 af 647, by BioLegend (1 mentions)
Cd45.2 af488, by BioLegend (1 mentions)
Cd11c percpcy5, by Cytek Biosciences (1 mentions)
Cd3ε pe cy7, by BioLegend (1 mentions)
Cd64 pe, by BioLegend (1 mentions)
Lsrfortessa flow cytometer, by BD (1 mentions)
Cd90.2 buv395, by BD (1 mentions)
Mhcii af700, by BioLegend (1 mentions)
Cytofix cytoperm kit, by BD (1 mentions)
2 protocols using cd8a buv 737
1
Multiparameter Flow Cytometry of Lymph Node T-cells
2
Skin and Lymph Node Immune Cell Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Single cell suspensions were obtained from uninfected skin and lymph nodes as previously described [13] (link). Intracellular CD207 staining was performed using BD Bioscience Cytofix/Cytoperm kit (BD Biosciences, San Jose, CA). For extracellular staining, antibodies were diluted in staining media (3% Calf Serum, 5mM EDTA, 0.04% Azide) with Fb block. The following antibodies were used for skin: CD45.2 AF488, CD64 PE, TCRβ PE-Dazzle 594, CD3ε PE-Cy7, Gr1 AF 647, MHCII AF 700, CD11b BV421, TCRγδ BV510 (all Biolegend, SanDiego, CA), CD11c PerCPCy5.5 (Tonbo Biosciences, San Diego, CA), CD90.2 BUV395, CD8a BUV737 (both BD Biosciences, San Jose, CA). For lymph node staining we used: CD45.2 AF488, CD64 PE, Gr1 AF 647, MHCII AF 700, CD11b BV421, (all Biolegend, SanDiego, CA), CD11c PerCPCy5.5 (Tonbo Biosciences, San Diego, CA). Sample data was acquired on an LSRFortessa flow cytometer (Becton Dickinson, Franklin Lakes, NJ), and analysed using FlowJo software (TreeStar, Ashland, OR).
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