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α cd8α

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α-CD8α is a monoclonal antibody that specifically binds to the CD8α subunit of the CD8 co-receptor on the surface of T cells and natural killer (NK) cells. It can be used for the identification and enumeration of CD8+ T cells and NK cells in flow cytometry applications.

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Lab products found in correlation

α cd4, by BioLegend (3 mentions) Facs lysis buffer, by BD (2 mentions) α cd40, by BioLegend (2 mentions) Collagenase type 4, by Thermo Fisher Scientific (2 mentions) α cd44, by BD (2 mentions) α cd45, by BioLegend (2 mentions) α cd69, by BioLegend (2 mentions) α cd103, by BioLegend (2 mentions) α ifnar1, by BioLegend (2 mentions) Gentlemacs, by Merck Group (2 mentions) Dnase 1, by Merck Group (2 mentions) Flow cytometry perm buffer, by Cytek Biosciences (1 mentions) Cd19 6d5, by BioLegend (1 mentions) Eomes dan11mag, by Thermo Fisher Scientific (1 mentions) Fixable viability dye 780, by Thermo Fisher Scientific (1 mentions) Cd44 im7, by Cytek Biosciences (1 mentions) Cd127 a7r34, by Cytek Biosciences (1 mentions) Klrg1 2f1 klrg1, by BioLegend (1 mentions) Cd45.1 a20, by BioLegend (1 mentions) Cd45.2 104, by BioLegend (1 mentions)

4 protocols using α cd8α

1

Characterization of CD8+ T Cell Subsets

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Cells were incubated with αCD16/32 (clone 2.4G2; hybridoma supernatant) and plated on U-bottom 96-well plates at ≤ 3 × 106 cells/well in complete RPMI. Where indicated, cells were stained with Kb-SIINFEKL tetramer APC (NIH tetramer core) at 37°C for 30 min in the presence of αCD8α (53–6.7; Biolegend). For live-dead and surface staining, cells were washed with media, and stained in media for 20 min at RT with Fixable Viability Dye 780 (eBioscience) and surface antibodies for CD19 (6D5, Biolegend), CD8α (53–6.7, Biolegend), CD44 (IM-7; Tonbo), CD127 (A7R34, Tonbo), KLRG1 (2F1/KLRG1, Biolegend), CD45.1 (A20, Biolegend), CD45.2 (104, Biolegend), CD122 (TM-b1, Biolegend). After wash with RPMI, cells were fixed and permeabilized for 45 min at RT in Foxp3/Transcription Factor 1× Fix/Perm solution (Tonbo), followed by wash with 1× Flow Cytometry Perm Buffer (Tonbo) and intracellular staining for 45 min at RT with intracellular antibodies, including TCF1 (C63D9; Cell Signaling Technology), FOXO1 (C29H4, Cell Signaling Technology), T-bet (4B10, Biolegend), EOMES (Dan11mag; eBioscience). The cells were washed twice and resuspended in 1× Flow Cytometry Perm Buffer (Tonbo). Flow cytometry data was acquired on a four-laser (405, 488, 561, 638 nm) CytoFlex S (Beckman Coulter) and analyzed using FlowJo software (BD Biosciences).
Ivanova D.L., Thompson S.B., Klarquist J., Harbell M.G., Kilgore A.M., Lasda E.L., Hesselberth J.R., Hunter C.A, & Kedl R.M. (2023). Vaccine adjuvant-elicited CD8+ T cell immunity is co-dependent on T-bet and FOXO1. Cell reports, 42(8), 112911.
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2

Comprehensive Immune Cell Profiling of Mouse Tissues

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At the designated times after infection, mice were euthanized, and tissues were perfused with PBS by cardiac injection. Lungs and/or dLN were harvested and single cell suspensions were created by enzymatic digestion with type IV collagenase (Fisher/Worthington) and DNase I (Sigma) and GentleMacs (Miltemyi) processing.
For flow cytometry staining, single cell homogenates were blocked in 2% rat serum for 10 min at room temperature, incubated with specified antibodies for 30 min on ice, fixed with BD FACS Lysis buffer for 10 min at room temperature, resuspended in PBS, and run on the LSRII (BD). Samples were analyzed using FlowJo software (BD).
The following anti-mouse antibodies conjugated to fluorochromes were utilized for flow cytometry: (company, clone) α-CD3ε (eBioscience/Invitrogen, 145–2C11), α-CD4 (Biolegend, GK1.5), α-CD8α (Biolegend, 53–6.7), α-CD11b (eBioscience/Invitrogen, M1/70), α-CD11c (eBioscience/Invitrogen, N418), α-CD19 (eBioscience/Invitrogen, 1D3), α-CD40 (Biolegend, HM40–3), α-CD44 (BD, IM7), α-CD45.1 (eBioscience/Invitrogen, A20), α-CD45.2 (Biolegend, 104), α-CD69 (Biolegend, H1.2F3), α-CD103 (Biolegend, 2E7), α-F4/80 (eBioscience/Invitrogen, BM8), α-IFNAR1 (Biolegend, MAR1–5A3), α-IFNγ (eBioscience/Invitrogen, XMG1.2), α-MHCII (eBioscience/Invitrogen, M5/114.15.2), α-TNFα (eBioscience/Invitrogen, MP6-XT22).
Hemann E.A., Green R., Turnbull J.B., Langlois R.A., Savan R, & Gale M J.r. (2019). Interferon-λ modulates dendritic cells to facilitate T cell immunity during infection with influenza A virus. Nature immunology, 20(8), 1035-1045.
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3

Mouse Cell Phenotype Analysis

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For analysis of mouse cell phenotypes, the following monoclonal antibodies were used: α-CD3 (clone: 145-2C11), α-CD4 (RM4-5), α-CD8α (53–6.7), α-CD25 (PC61), α-CD44 (IM7), α-CD62L (MEL14), α-CD90.1 (OX-7), α-CD40L (MR-1), α-CD11c (HL3), α-CD11b (M1/70), α-Gr1 (RB6-8C5), α-Ly6C (HK1.4), α-Ly6G (1A8), α-F4/80 (BM8), α-IFN-γ (XMG1.2), α-IL-17A (TC11-18H10), α-GM-CSF (MP1-22E9; all from BioLegend) and α-IL-22 (AM22-3; provided by J.C. Renauld). For staining of lipid rafts, biotin-conjugated cholera toxin subunit B (Molecular Probes) was used together with fluorochrome-conjugated streptavidin (BioLegend). Dead cells were always excluded by staining with 7-AAD (BioLegend) or 4,6-diamidino-2-phenylindole (Sigma). All antibodies were used at 1 μg per 106 cells. For intracellular cytokine staining, cells were stimulated for 5 h with phorbol 12-myristate 13-acetate (10−7 M; Sigma) and ionomycin (1 μg ml−1; Sigma), in the presence of brefeldin A (10 μg ml−1; Sigma) for the last 3 h of culture. Cells were fixed with 4% (wt/vol) paraformaldehyde and permeabilized with 0.5% (wt/vol) saponin (Sigma). Eight-colour staining was performed with the appropriate combinations of antibodies conjugated to fluorochromes. Samples were acquired on a FACSCanto II (BD Biosciences) and analysed with FlowJo software (TreeStar) and when necessary using the Boolean gating strategy65 (link).
Ronchi F., Basso C., Preite S., Reboldi A., Baumjohann D., Perlini L., Lanzavecchia A, & Sallusto F. (2016). Experimental priming of encephalitogenic Th1/Th17 cells requires pertussis toxin-driven IL-1β production by myeloid cells. Nature Communications, 7, 11541.
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4

Comprehensive Immune Cell Profiling of Mouse Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols
At the designated times after infection, mice were euthanized, and tissues were perfused with PBS by cardiac injection. Lungs and/or dLN were harvested and single cell suspensions were created by enzymatic digestion with type IV collagenase (Fisher/Worthington) and DNase I (Sigma) and GentleMacs (Miltemyi) processing.
For flow cytometry staining, single cell homogenates were blocked in 2% rat serum for 10 min at room temperature, incubated with specified antibodies for 30 min on ice, fixed with BD FACS Lysis buffer for 10 min at room temperature, resuspended in PBS, and run on the LSRII (BD). Samples were analyzed using FlowJo software (BD).
The following anti-mouse antibodies conjugated to fluorochromes were utilized for flow cytometry: (company, clone) α-CD3ε (eBioscience/Invitrogen, 145–2C11), α-CD4 (Biolegend, GK1.5), α-CD8α (Biolegend, 53–6.7), α-CD11b (eBioscience/Invitrogen, M1/70), α-CD11c (eBioscience/Invitrogen, N418), α-CD19 (eBioscience/Invitrogen, 1D3), α-CD40 (Biolegend, HM40–3), α-CD44 (BD, IM7), α-CD45.1 (eBioscience/Invitrogen, A20), α-CD45.2 (Biolegend, 104), α-CD69 (Biolegend, H1.2F3), α-CD103 (Biolegend, 2E7), α-F4/80 (eBioscience/Invitrogen, BM8), α-IFNAR1 (Biolegend, MAR1–5A3), α-IFNγ (eBioscience/Invitrogen, XMG1.2), α-MHCII (eBioscience/Invitrogen, M5/114.15.2), α-TNFα (eBioscience/Invitrogen, MP6-XT22).
Hemann E.A., Green R., Turnbull J.B., Langlois R.A., Savan R, & Gale M J.r. (2019). Interferon-λ modulates dendritic cells to facilitate T cell immunity during infection with influenza A virus. Nature immunology, 20(8), 1035-1045.
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