Cd3 bv421 clone 17a2

Manufactured by BioLegend

Sourced in United States

CD3-BV421 (clone 17A2) is a fluorochrome-conjugated antibody that binds to the CD3 surface antigen. CD3 is a complex of membrane-bound proteins that is essential for the activation of T cells. This antibody can be used to identify and enumerate T cells in flow cytometry applications.

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Lab products found in correlation

Tim3 apc clone rmt3 23, by BioLegend (1 mentions) Ficoll paque plus density gradient media, by GE Healthcare (1 mentions) Clone gk1, by BioLegend (1 mentions) Cd4 apc cy7, by BioLegend (1 mentions) Kaluza software, by Beckman Coulter (1 mentions) Gallios flow cytometer, by Beckman Coulter (1 mentions) Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Accucheck counting beads, by Thermo Fisher Scientific (1 mentions) Cd4 fitc clone rm4 4, by Thermo Fisher Scientific (1 mentions) Lsrii flow cytometer, by BD (1 mentions)

Close spelling variants of this product

CD3 (17A2; (30 citations) CD3 (clone 17A2, (24 citations) Clone 17A2 (20 citations) CD3-BV421 (20 citations) Anti-CD3-BV421 (clone 17A2, (3 citations)

2 protocols using cd3 bv421 clone 17a2

Characterizing Tumor-Infiltrating Lymphocytes

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End-point tumours (150 mm²) from VP60 and surv.VLP-SS-MUC1 treated mice were resected for tumour infiltrating lymphocyte (TIL) analysis. Five tumours per group were fixed in formalin for CD3 immunohistochemistry (Histology Unit, University of Otago). Images were taken with Aperio CS2 and analysed using ImageJ software (National Institutes of Health). The other five tumours per group were processed into single-cell suspensions by cutting them into small pieces using a scalpel, then sequentially passing through 100 and 70 μm cell strainers, followed by density gradient centrifugation (Ficoll-Paque PLUS density gradient media, GE Healthcare). TILs were stained with Zombie-Yellow Live/Dead Stain, treated with CD16/CD32 Fc blocking antibody, and then stained with PD-1-FITC (clone 29F.1A12, BioLegend, San Diego, CA, USA), LAG3-PE (clone C9B7W, BD), CD127-PE/CF594 (clone SB/199, BioLegend), CD39-PE/Cy7 (clone Duha59, BioLegend), TIM3-APC (clone RMT3-23, BioLegend), CD8-AF700 (clone 53-6.7, BioLegend), CD4-APC/Cy7 (clone GK1.5, BioLegend) and CD3-BV421 (clone 17A2, BioLegend). Fluorescence was measured using a Gallios flow cytometer and analysed using Kaluza software (Beckman Coulter, Brea, CA, USA).

Campbell K., Young V.L., Donaldson B.C., Woodall M.J., Shields N.J., Walker G.F., Ward V.K, & Young S.L. (2021). Delivering Two Tumour Antigens Survivin and Mucin-1 on Virus-Like Particles Enhances Anti-Tumour Immune Responses. Vaccines, 9(5), 463.

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Multiparameter Flow Cytometry of Immune Cells

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For flow cytometric analyzes, recovered immune cells from processed blood and organs were first stained for surface markers using the following anti-mouse antibodies: CD3 BV421 (clone 17A2, BioLegend), CD4 FITC (clone RM4-4, eBioscience), CD8a BV605 (clone 53–6.7, BioLegend), and/or CD44 AF700 (clone IM7, BioLegend). Live/dead cell discrimination was done using LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (Thermo Fisher Scientific). Cells were then fixed followed by staining with anti-mouse FOXP3 AF700 or APC (clone FJK-16s, eBioscience). For blood samples, 25 uL of AccuCheck Counting Beads (Thermo Fisher Scientific) were added to determine absolute cell counts. Samples were acquired on an LSR II Flow Cytometer (Becton Dickinson). Flow cytometry data were analyzed using FlowJo software (FlowJo, LLC). Lymphocytes were gated on using side vs. forward scatter plots followed by doublet and live/dead discrimination. CD8+ and CD4+ T cells were then gated on live CD3+ T cells. Regulatory T cells (Treg) and effector CD8+ T cells (CD8_eff) were defined as CD4+ FOXP3+ and CD8+ CD44^hi cells, respectively. Fluorescence Minus One (FMO) controls were used to guide FOXP3 and CD44 gating strategies. Absolute cell counts were calculated using the counting beads according to the manufacturer’s protocol.

Maxwell R., Luksik A.S., Garzon-Muvdi T., Hung A.L., Kim E.S., Wu A., Xia Y., Belcaid Z., Gorelick N., Choi J., Theodros D., Jackson C.M., Mathios D., Ye X., Tran P.T., Redmond K.J., Brem H., Pardoll D.M., Kleinberg L.R, & Lim M. (2018). Contrasting impact of corticosteroids on anti-PD-1 immunotherapy efficacy for tumor histologies located within or outside the central nervous system. Oncoimmunology, 7(12), e1500108.

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