Cd8 brilliant violet 421

Manufactured by BioLegend

The CD8-Brilliant Violet 421 is a fluorescently-labeled antibody that binds to the CD8 protein, which is expressed on the surface of certain T cells. This product can be used for the identification and enumeration of CD8+ T cells in flow cytometry applications.

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Brilliant Violet 421 (66 citations)

4 protocols using cd8 brilliant violet 421

Quantifying Human Cell Populations

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Humanized mice-derived samples were analyzed for their human cell distribution by flow-cytometry using the following antibodies from Biolegend: CD19-APC (#302212), CD3-PE (#300308), CD4-PECy7 (#300512), CD8-Brilliant Violet 421 (#301036). CD38-PerCPCy5.5 (#303521) and HLA-DR-FITC (#307604). The pan-human marker CD45-Krome Orange, was obtained from Beckman Coulter (PN96416).

Rochat M.A., Schlaepfer E., Kuster S.P., Li D., Audige A., Ivic S., Fahrny A, & Speck R.F. (2018). Monitoring HIV DNA and cellular activation markers in HIV-infected humanized mice under cART. Virology Journal, 15, 191.

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Flow Cytometry for T Cell Analysis

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Fluorescence conjugated anti-mouse antibodies CD4-PerCP-Cy5.5 (GK1.5) and CD8-Brilliant Violet 421 (53–6.7) were from Biolegend (San Diego, CA). FoxP3-FITC (FJK-16s) was from eBioscience (San Diego, CA). H-2D^b tetramers loaded with the E7 peptide (RAHYNIVTF) was kindly provided by the National Institute of Allergy and Infectious Diseases Tetramer Core Facility and the National Institutes of Health AIDS Research and Reference Reagent Program. CountBright^™ absolute counting beads were from Life Technologies (Grand Island, NY).

Chen Z., Ozbun L., Chong N., Wallecha A., Berzofsky J.A, & Khleif S.N. (2014). Episomal expression of truncated listeriolysin O in LmddA-LLO-E7 vaccine enhances antitumor efficacy by preferentially inducing expansions of CD4+FoxP3− and CD8+ T cells. Cancer immunology research, 2(9), 911-922.

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Intranasal Ragweed Pollen Immunization in Mice

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Wildtype C57Bl/6 mice were lightly anaesthetised with isoflurane and immunised with 50 μl of short ragweed pollen (100 μg/dose of protein, Greer Laboratories) intranasally for four consecutive days. Mice were euthanized 24 hours after final administration and tissues were collected for analysis. Lung tissues were chopped into small pieces then incubated with 720 μg/ml collagenase D (Amersham, Bucks) for 1 hour. Lung and mediastinal lymph node tissues were mechanically passed through 70 μm cell strainers to obtain single cell suspensions. Tissue cell suspensions were incubated with anti-Fc receptor blocking antibody (anti-CD16/32, eBioscience, 14-0161-85), then stained with the following antibody panel: CD19-PerCP-Cy5.5 (eBioscience, 45-0193-82), ICOS-Alexa Fluor 647 (Biolegend, 313516), CD4-Alexa Fluor 700 (eBioscience, 56-0041-80), Fixable Viability Dye eFluor780 (eBioscience, 65-0865-14), CD8-Brilliant Violet 421 (Biolegend, 100738) and Lineage-PeCy7 (CD3 (eBioscience, 25-0031-82), CD11b (eBioscience, 25-0112-82), CD11c (eBioscience, 25-0114-82), FcεR1 (eBioscience, 25-5898-82), Gr-1 (eBioscience, 25-5931-82), NK1.1 (eBioscience, 25-5941-82), Ter-119 (eBioscience, 25-5921-82)).

Beale J., Jayaraman A., Jackson D.J., Macintyre J.D., Edwards M.R., Walton R.P., Zhu J., Man Ching Y., Shamji B., Edwards M., Westwick J., Cousins D.J., Yi Hwang Y., McKenzie A., Johnston S.L, & Bartlett N.W. (2014). Rhinovirus induced IL-25 in asthma exacerbation drives type-2 immunity and allergic pulmonary inflammation. Science translational medicine, 6(256), 256ra134.

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Profiling Immune Cells by Flow Cytometry

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Monoclonal antibodies specific for human CD3-PeCy7, CD3-APC-Cy7 (Clone HIT3a), CD4-Alexa Fluor 488 (Clone OKT4), CD8-APC-Cy7, CD8-Brilliant Violet 421 (Clone RPA-T8), IgG1-PE (Clone HP6017), IgG1 Isotype-PE (Clone MOPC−21), 41BB-PE (Clone 4B4–1), PD−1-PeCy7, PD−1-Brilliant Violet 421 (Clone EH12.2H7), CD69-APC-Cy7 (Clone FN50), IFNγ-APC-Cy7 (Clone 4S.B3), TNFα-Alexa Fluor 488 (Clone Mab11), granzyme B-APC (Clone GB−11), perforin-PerCPCy5.5 (Clone B-D48), CEA-APC (Clone ASL−32), PD-L1-Brilliant Violet 421 (Clone 29E.2A3), TruStain fcX™ (Clone 93) and viability dye Zombie Aqua (ref 423,101), were purchased from Biolegend. Samples were analyzed in a FACSCanto II cytometer (BD Bioscience) and data were analyzed using FlowJo version X.0.7 (Tree Star, Inc.).

Lopez E., Hidalgo S., Roa E., Gómez J., Hermansen Truan C., Sanders E., Carrasco C., Pacheco R., Salazar-Onfray F., Varas-Godoy M., Borgna V, & Lladser A. (2023). Preclinical evaluation of chimeric antigen receptor T cells targeting the carcinoembryonic antigen as a potential immunotherapy for gallbladder cancer. Oncoimmunology, 12(1), 2225291.

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