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Anti cd8 apc cy7 clone 53 6.7

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Anti-CD8-APC-Cy7 (Clone 53-6.7) is a fluorochrome-conjugated monoclonal antibody that binds to the CD8 surface marker. This antibody is commonly used in flow cytometry applications for the detection and analysis of CD8-positive cells.

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Lab products found in correlation

Cell light apollo488 stain kit, by RiboBio (1 mentions) Fitc annexin 5 apoptosis detection kit, by BD (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)

2 protocols using anti cd8 apc cy7 clone 53 6.7

1

Tracking Proliferation and Apoptosis in Transplanted Mice

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Recipient mice were pulsed intraperitoneally with 0.1 mg of 5-ethynyl-2′-deoxyuridine (EDU, RIBOBIO) in PBS 10 days after transplantation. 24 h later, spleen and LN cells were isolated and single-cell suspensions were prepared for EDU detection using Cell-Light™ Apollo®488 Stain Kit (RIBOBIO) according to the manufacturer’s instructions. Then, cells were stained for cell surface markers with anti-CD8-APC-Cy7 (Clone 53-6.7), CD44-V450 (Clone IM7), and CD62L-APC Abs (Clone MEL-14) (all from BD Biosciences). To detect cell apoptosis, cells were stained for the same surface markers and then Annexin V-FITC according to the protocol of Annexin V-FITC Apoptosis Detection Kit (BD Pharmingen). Cells finally were analyzed via a flow cytometer (FACSCalibur).
Qiu F., Liu H., Liang C.L., Nie G.D, & Dai Z. (2017). A New Immunosuppressive Molecule Emodin Induces both CD4+FoxP3+ and CD8+CD122+ Regulatory T Cells and Suppresses Murine Allograft Rejection. Frontiers in Immunology, 8, 1519.
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2

Cytolytic Assay for CAR T Cells

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A flow cytometry based assay was used to measure the cytolytic activity of the CAR T cells. 105 DR1-CII or DR1-HA CAR T cells were co-cultured in 300 μl RPMI medium in a 96 well microtiter plate with 104 cloned DR1-restricted T cells for a 10:1 effector:target (E:T) ratio. Targets consisted of T cell hybridomas specific for either the CII peptide (GIAGFKGEQGPKGEP) or the HA peptide (PKYVKQNTLKLAT). After 4 hr at 37° C, cells were collected from the wells and stained with anti-CD8-APC-Cy7 (clone 53-6.7, BD), anti-BV14-FITC (clone 14.2, BD), anti-CD4-PerCP-Cy5.5 (clone RM4-5, Tonbo), anti-BV8-PE (clone F23.1, BD), anti-HLA-DR-Pacific Orange (clone LB 3.1), and DAPI (Sigma) was added just prior to analysis for exclusion of dead cells. Data analyses were focused on live cells (DAPI negative), HLA-DR1 negative, CD8 negative, and CD4+/BV14+ (HA specific) or CD4+/BV8+ (CII-specific) T cells.
Whittington K.B., Prislovsky A., Beaty J., Albritton L., Radic M, & Rosloniec E.F. (2021). CD8+ T cells expressing an HLA-DR1 chimeric antigen receptor (CAR) target autoimmune CD4+ T cells in an antigen specific manner and inhibit the development of autoimmune arthritis. Journal of immunology (Baltimore, Md. : 1950), 208(1), 16-26.
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