Bv421 mouse anti human cd8

Manufactured by BD

Sourced in United States

The BV421 mouse anti-human CD8 is a fluorescently labeled monoclonal antibody that binds to the CD8 receptor on the surface of human T cells. It is used for the identification and quantification of CD8+ T cells in flow cytometry applications.

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4 protocols using bv421 mouse anti human cd8

Optimizing HIV-1 Virus Detection Assay

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Native CAB and CAB LAP (200 mg/mL) were graciously obtained from ViiV Healthcare (Research Triangle Park, NC, USA). Myristoyl chloride, poloxamer 407 (P407), N,N-diisopropylethylamine (DIEA), dimethylformamide (DMF), 1-octanal, ciprofloxacin, paraformaldehyde (PFA), and 3,3′-diaminobenzidine (DAB) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s Modified Eagle’s Medium (DMEM) was purchased from Corning Life Sciences (Tewksbury, MA, USA). Heat-inactivated pooled human serum was purchased from Innovative Biologics (Herndon, VA, USA). Cell counting kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies, Inc. (Rockville, MD, USA). Gentamicin, HPLC grade acetonitrile (ACN), HPLC grade methanol, and Optima grade LC/MS water was purchased from ThermoFisher Scientific (Waltham, MA, USA). FITC mouse anti-human CD45, Alexa Fluor 700 mouse anti-human CD3, APC mouse anti-human CD4, and BV421 mouse anti-human CD8 were purchased from BD Biosciences (San Jose, CA, USA). Monoclonal mouse anti-human HIV-1p24 (clone Kal-1), monoclonal mouse anti-human leukocyte antigen (HLA-DR; clone CR3/43), and the polymer-based HRP-conjugated anti-mouse EnVision + secondary were purchased from Dako (Carpinteria, CA, USA).

Zhou T., Su H., Dash P., Lin Z., Shetty B.L., Kocher T., Szlachetka A., Lamberty B., Fox H.S., Poluektova L., Gorantla S., McMillan J., Gautam N., Mosley R.L., Alnouti Y., Edagwa B, & Gendelman H.E. (2017). Creation of a nanoformulated cabotegravir prodrug with improved antiretroviral profiles. Biomaterials, 151, 53-65.

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Optimizing HIV-1 Virus Detection Assay

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Characterization of Nanoformulated HIV Therapy

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DTG was a generous gift from ViiV Healthcare (Research Triangle Park, NC). Pyridine, dimethylformamide (DMF), N,N-diisopropylethylamine (DIEA), myristoyl chloride, poloxamer 407 (P407), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, ciprofloxacin, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), paraformaldehyde (PFA), and 3,3’-diaminobenzidine (DAB) were purchased from Sigma-Aldrich (St. Louis, MO). Diethyl ether, cell culture grade water (endotoxin-free), gentamicin, acetonitrile (ACN), methanol, KH₂PO₄, bovine serum albumin (BSA), Triton X-100, LC-MS-grade water, and TRIzol reagent were purchased from Fisher Scientific (Hampton, NH). FITC mouse anti-human CD45, Alexa Fluor 700 mouse anti-human CD3, APC mouse anti-human CD4, BV421 mouse anti-human CD8, PE mouse anti-human CD14, and PE-Cy5 mouse anti-human CD19 were purchased from BD Biosciences (San Jose, CA). Monoclonal mouse anti-human HIV-1p24 (clone Kal-1), monoclonal mouse anti-human leukocyte antigen (HLA-DP/DQ/DR; clone CR3/43), and the polymer-based HRP-conjugated anti-mouse EnVision+ secondary were purchased from Dako (Carpinteria, CA). Heat-inactivated pooled human serum was purchased from Innovative Biologics (Herndon, VA). Dulbecco’s Modification of Eagle’s Medium (DMEM) was purchased from Corning Life Sciences (Tewksbury, MA).

Sillman B., Bade A.N., Dash P.K., Bhargavan B., Kocher T., Mathews S., Su H., Kanmogne G.D., Poluektova L.Y., Gorantla S., McMillan J., Gautam N., Alnouti Y., Edagwa B, & Gendelman H.E. (2018). Creation of a long-acting nanoformulated dolutegravir. Nature Communications, 9, 443.

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Cytotoxicity Assay for CAR-T Cell Evaluation

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Cytotoxicity assays were carried out using the flow cytometer (Except for Fig. 1a). Briefly, CAR-T cells were co-cultrured with zipFv and target cells for 24 h at 37 °C. Prior to flow cytometry, control samples containing only the target cells were used to set a flow cytometry gate for intact target cells based on forward and side scatter patterns that had been previously confirmed to exclude dead cells (Supplementary Fig. 3e). Also, fluorescent proteins and staining CD19 (anti-Human CD19 PE-Cy7 (1:200 dilution, Tonbo Bioscience, clone HIB19)) were used to further identify the target cells, or staining CD8 (BV421 Mouse Anti-Human CD8 (1:200 dilution, BD)) was used to exclude CAR-T cells This gate was applied to all other samples. Live target cell number was calculated and target cell cytotoxicity was calculated using the following formula: Cytotoxicity = 100 × [(Total live target cell number − number of remaining live cells after lysis)/(Total live target cell number)].

Cho J.H., Okuma A., Sofjan K., Lee S., Collins J.J, & Wong W.W. (2021). Engineering advanced logic and distributed computing in human CAR immune cells. Nature Communications, 12, 792.

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