Anti cd8a af700

Manufactured by BioLegend

Anti-CD8a AF700 is a fluorescently-labeled antibody that binds to the CD8a surface antigen. It is designed for use in flow cytometry applications.

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Anti-CD8a (26 citations) CD8a-AF700 (4 citations)

3 protocols using anti cd8a af700

Multiparameter Immune Cell Profiling

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Anti–CD3ε-PerCP (100326), anti–CD8a-AF700 (100730), anti–CD8a-APC (100712), anti–PD-1-PE/DZ594 (109116), anti–Ki67-FITC (652410), anti–TCF1-PE (655208), anti–IFN-γ-BV421 (505830), anti–IFN-γ-PerCp/Cy5.5 (505822), anti–TNF-α-APC/Cy7 (506344), anti–TIM-3-PE/Cy7 (119716), anti–CD45-BV510 (103138), anti–Ly108 (Slamf6)-PE (134606), anti–PD-1-FITC (135214), anti–CD45.1-PE (110708), anti–CD45.2-AF488 (109816), purified anti-mouse CD3ε (100340), and purified anti-mouse CD28 (102116) were obtained from Biolegend. Anti–CD8-FITC (D271-4) and tetramer-SIINFEKL-APC (TS-5001-2c) were purchased from MBL. Cell Stimulation Cocktail Plus Protein Transport Inhibitors and FOXP3/TF Staining Buffer Set were bought from eBioscience. Naive CD8a⁺ T Cell Isolation and Tumor Dissociation Kits were obtained from Miltenyi Biotec.

Li X., Li Y., Dong L., Chang Y., Zhang X., Wang C., Chen M., Bo X., Chen H., Han W, & Nie J. (2023). Decitabine priming increases anti–PD-1 antitumor efficacy by promoting CD8+ progenitor exhausted T cell expansion in tumor models. The Journal of Clinical Investigation, 133(7), e165673.

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Tumor Immune Profiling by Flow Cytometry

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Tumors were collected 3 days after last radiation fraction then were dissociated in DMEM supplemented with 1 mg/ml Collagenase IV (Solarbio) and 1 μl/ml DNase I (Solarbio) at 37 °C for one hour. Single-cell suspensions were collected after filtering digested tissues with 40-um filter and washed with PBS supplemented with 2% FBS. Subsequently, cell surface and intracellular markers were stained with the following fluorochrome-conjugated antibodies: anti-CD45 BV421 (Cat#: 103,134), anti-CD11b PE (Cat#: 101,207), anti-F4/80 AF700 (Cat#: 123,129), anti-CD86 FITC (Cat#: 105,109), anti-CD206 APC (Cat#: 141,708), anti-CD4 BV510 (Cat#: 100,553), anti-CD8a AF700 (Cat#: 100,729) from Biolegend. For surface staining, all samples were stained with antibodies at 4 °C for 30 min; for intracellular staining, cells were fixed and permeabilized with BD Cytofix/Cytoperm (Cat#: 554,714) then stained according to the manufacturer’s instructions. Analysis of stained cells was performed using a CytoFlex cytometer (Beckman Coulter) and CytExpert software.

Yang H., Lei Z., He J., Zhang L., Lai T., Zhou L., Wang N., Tang Z., Sui J, & Wu Y. (2024). Single-cell RNA sequencing reveals recruitment of the M2-like CCL8high macrophages in Lewis lung carcinoma-bearing mice following hypofractionated radiotherapy. Journal of Translational Medicine, 22, 306.

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OP9-DL1 Co-culture for T-cell Development

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OP9 bone marrow stromal cells expressing the Notch ligand DL-1 (OP9-DL1) were kindly provided by Dr. Juan Carlos Zúñiga-Pflücker (University of Toronto). The OP9-DL1 cells were cultured and maintained in αMEM medium, supplemented with 10% FBS (Gibco), penicillin (100 µg/mL) and streptomycin (100 U/mL) (Invitrogen). For thymocytes co-cultures, the sorted DN2 or DN3 cells were plated onto confluent OP9-DL1 monolayers (70-80% confluent) with addition of 5 ng/ml recombinant murine interleukin-7 (PeproTech) and 5 ng/ml Flt3L (PeproTech). Cells were harvested at different time points, filtered through 40µm cell strainer, and stained with antibodies including ani-CD45 V450 (Tonbo, 75-0451-U100), anti-CD4 BV650 (Biolegend, 100546), anti-CD8a AF700 (Biolegend, 100730), anti-CD44 FITC (Biolegend, 103006), or anti-CD25 APC (Biolegend, 101910), followed by staining with Annexin V PE (Biolegend, 640908) and DAPI (4,6-diamidino-2-phenylindole, Invitrogen, D1306). EL4 cells (ATCC-TIB39) were cultured in DMEM with 10% FBS (Gibco), penicillin (100 µg/mL) and streptomycin (100 U/mL) (Invitrogen).

Liu X., Yu J., Xu L., Umphred-Wilson K., Peng F., Ding Y., Barton B.M., Lv X., Zhao M.Y., Sun S., Hong Y., Qi L., Adoro S., & Chen X. (2021). Notch-Induced Endoplasmic Reticulum-Associated Degradation Governs Thymocyte β−Selection.

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