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Cd8a af700

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CD8a-AF700 is a fluorescently labeled antibody product from BioLegend. It is designed to detect the CD8a surface marker on T cells. The antibody is conjugated to the Alexa Fluor 700 fluorescent dye.

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Lab products found in correlation

Cytofix cytoperm, by BD (2 mentions) Siglecf pe, by BD (2 mentions) Cd4 pe cy7, by BioLegend (2 mentions) Mhcii fitc, by BioLegend (2 mentions) B220 fitc, by BioLegend (2 mentions) Perm buffer, by Thermo Fisher Scientific (2 mentions) Cell stimulation cocktail with protein inhibitors, by Thermo Fisher Scientific (2 mentions) Cd45 sb600, by BioLegend (2 mentions) Tcrb apc cy7, by BioLegend (2 mentions) Cd8b pe, by BioLegend (2 mentions) Ifng percp cy5, by BioLegend (2 mentions) Tnfa apc, by BioLegend (2 mentions) Tcrg fitc, by BioLegend (2 mentions) Mhcii apc e780, by BioLegend (2 mentions) Il 17a bv421, by BioLegend (2 mentions) Cd11b percp cy5, by BioLegend (2 mentions) Ly6g pe cy7, by BD (2 mentions) Cd3e fitc, by BioLegend (2 mentions) Cd11c af700, by BioLegend (2 mentions) Cd64 apc, by BioLegend (2 mentions)

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Anti-CD8a AF700 (3 citations)

4 protocols using cd8a af700

1

Multiparametric Flow Cytometry of Tumor Cells

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Single-cell suspensions of tumors were prepared, and flow cytometry was carried out as mentioned earlier [45 (link), 46 (link)]. Cells were stained with the following antibodies obtained from BioLegend: CD16/32 (clone 93, catalog No. 103132), Zombie UV™ Fixable Viability Kit (catalog No. 100752), CD45-PerCP/Cy5.5 (clone 30-F11, catalog No. 103132), CD45-APC/Cy7 (clone 30-F11, catalog No. 103116), CD11b-FITC (clone M1/70, catalog No. 101206), SIRPα-PE (clone BM8, catalog No. 123122), Gr-1-PerCP/Cy5.5 (clone RB6-8C5, catalog No. 108428), F4/80-AF647 (clone N418, catalog No. 117318),CD11c-PE/Cy7 (clone N418, catalog No. 117318), I-A/I-E-AF700 (clone M5/114.15.2, catalog No. 107622), CD86-BV421 (clone GL-1, catalog No. 105032), CD206-BV605 (clone C068C2, catalog No. 141721), CD8a-AF700 (clone 53-6.7, catalog No. 100730), CD8-BV510 (clone 53-6.7, catalog No. 100752), TNF-α-BV421 (clone MP6-XT22, catalog No. 506328), IFN-γ-PE (clone XMG1.2, catalog No. 505808), CD39-PE/Cy7 (clone Duha59, catalog No. 143806), Tim-3-APC/Fire™ 750 (clone RMT3-23, catalog No. 119738), PD-1-BV605 (clone 29 F.1A12, catalog No. 135220). The next antibodies were obtained through eBioscience: Mouse Regulatory T Cell Staining Kit #2 (clone FJK-16s, catalog No. 88-8118-40). FlowJo program (Tree Star Inc.) was utilized to examine the findings that were acquired through a BD LSRFortessa Flow Cytometer.
Ji K., Zhang Y., Jiang S., Sun L., Zhang B., Hu D., Wang J., Zhao L., Wang P, & Tao Z. (2023). SIRPα blockade improves the antitumor immunity of radiotherapy in colorectal cancer. Cell Death Discovery, 9, 180.
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2

Comprehensive Immune Cell Profiling

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Intestinal and mesenteric lymph node immune cell populations were characterised by flow cytometry. For intracellular cytokine staining, cells were incubated in T cell media (RPMI, 10% FBS, 1 mM sodium pyruvate, 2mM GLUTamax, 1X non-essential amino acids, 0.1 mM 2-β-mercaptoethanol, 10 mM HEPES, 1%Penicillin/Streptomycin) + 1X Cell Stimulation Cocktail with Protein Inhibitors (eBioscience) for 3 hours at 37°C before staining. Non-viable cells were stained using Live/Dead Fixable Aqua. Cells were permeabilised with CytoFix/CytoPerm (BD) followed by intracellular staining in PermBuffer (eBioscience).
All antibodies were purchased from eBioscience unless otherwise indicated. Antibodies used were CD45-SB600, TCRb-APC-Cy7, MHCII-FITC, CD4-PE-Cy7, CD8a-AF700, CD8b-PE, IFNg-PerCP-Cy5.5, TNFa-APC, IL-17A-bv421 (BioLegend), CD3e-FITC, TCRg-FITC, B220-FITC, MHCII-APC-e780, CD11b-PerCP-Cy5.5, CD11c-AF700, CD64-APC (BioLegend), SiglecF-PE (BD), Ly6G-Pe-Cy7 (BD) and F4/80-e450. Sample data were acquired with an Attune NxT flow cytometer coupled with an Attune CytKick Max autosampler. Data were analysed using FlowJo v10. Antibody details and concentrations are included in Supplementary Table 1.
Forster S.C., Clare S., Beresford-Jones B.S., Harcourt K., Notley G., Stares M.D., Kumar N., Soderholm A.T., Adoum A., Wong H., Morón B., Brandt C., Dougan G., Adams D.J., Maloy K.J., Pedicord V.A, & Lawley T.D. (2022). Identification of gut microbial species linked with disease variability in a widely used mouse model of colitis. Nature Microbiology, 7(4), 590-599.
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3

Immune Cell Profiling in Intestine and Lymph Nodes

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Intestinal and mesenteric lymph node immune cell populations were characterized by flow cytometry. For intracellular cytokine staining, cells were incubated in T cell medium (RPMI, 10% FBS, 1 mM sodium pyruvate, 2 mM GLUTamax, 1× nonessential amino acids, 0.1 mM 2-β-mercaptoethanol, 10 mM HEPES, 1% penicillin/streptomycin) + 1× Cell Stimulation Cocktail with Protein Inhibitors (eBioscience) for 3 h at 37 °C before staining. Nonviable cells were stained using Live/Dead Fixable Aqua. Cells were permeabilized with CytoFix/CytoPerm (BD), followed by intracellular staining in PermBuffer (eBioscience).
All antibodies were purchased from eBioscience unless otherwise indicated. Antibodies used were CD45-SB600, TCRb-APC-Cy7, MHCII-FITC, CD4-PE-Cy7, CD8a-AF700, CD8b-PE, IFNg-PerCP-Cy5.5, TNFa-APC, IL-17A-bv421 (BioLegend), CD3e-FITC, TCRg-FITC, B220-FITC, MHCII-APC-e780, CD11b-PerCP-Cy5.5, CD11c-AF700, CD64-APC (BioLegend), SiglecF-PE (BD), Ly6G-Pe-Cy7 (BD) and F4/80-e450. Sample data were acquired with an Attune NxT flow cytometer coupled with an Attune CytKick Max autosampler. Data were analyzed using FlowJo v.10. Antibody details and concentrations are included in Supplementary Table 1.
Forster S.C., Clare S., Beresford-Jones B.S., Harcourt K., Notley G., Stares M.D., Kumar N., Soderholm A.T., Adoum A., Wong H., Morón B., Brandt C., Dougan G., Adams D.J., Maloy K.J., Pedicord V.A, & Lawley T.D. (2022). Identification of gut microbial species linked with disease variability in a widely used mouse model of colitis. Nature Microbiology, 7(4), 590-599.
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4

Multiparameter Flow Cytometry Analysis

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Single cells were incubated with Human TruStain FcX (BioLegend) to block Fc receptors. These were then stained with Ghost Dye™ Red 780 Viability Dye (Cell Signaling Technology) to identify dead cells and the following fluorochrome-conjugated anti-mouse monoclonal antibodies (mAbs): CD103-PerCP/Cy5.5 (Ber-ACT8), CTLA-4- PerCP/Cy5.5 (BNI3), Granzyme B-FITC (QA16A02), PD-1-PE (EH12.2H7), CD27- PE/Cy7 (LG.3A10), CD3- PE/Cy7 (HIT3a), CD8a-AF700 (RPA-T8), CD4-APC (OKT4), CD127-BV711 (A019D5), Ki-67-BV605 (Ki-67), CD45RA-BV605 (HI100), CCR7-BV510 (G043H7), CD69-BV421 (FN50), and IFNγ-BV421 (4S.B3) (all from BioLegend). Data were acquired on a LSRFortessa X-20 (BD) and analyzed with FlowJo software.
Miyabe C., Dong Y., Ikeda T., Takahashi K., Miyabe Y, & Kawakami T. (2021). Immune checkpoint molecule expression is altered in the skin and peripheral blood in vasculitis. Scientific Reports, 11, 20019.
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