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Multicolor flow cytometric analysis of TCRαβ expression on human peripheral blood lymphocytes. Whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD19 antibody (Cat. No. 659477), FITC Mouse Anti-Human CD3 antibody (Cat. No. 555916/555332/561807/561806) and either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat No. 566928; Left Plots) or BD Horizon™ R718 Mouse Anti-Human TCRαβ antibody (Cat. No. 567676/567677; Right Plots). Erythrocytes were lysed with BD FACSLyse™ Lysing Buffer (Cat. No. 349202). Flow cytometric analysis was performed using a LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ software. Upper Plots: Bivariate pseudocolor density plots showing the correlated expression of CD19 versus TCRαβ (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Bottom Plots: Bivariate pseudocolor density plots showing the correlated expression of CD3 versus TCRαβ (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.
BD Horizon™ R718 Mouse Anti-Human TCRαβ
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
The IP26 monoclonal antibody specifically recognizes a monomorphic determinant on the αβ T-cell receptor (TCRαβ). The TCRαβ is a disulfide-linked 80 kDa heterodimer consisting of a 44 kDa α chain and a 37 kDa β chain. The TCRαβ is normally expressed on 50-70% of thymocytes and on a large fraction of mature T cells including greater than 95% of peripheral blood CD3+ T cells. The TCRαβ serves as a receptor for peptide antigens bound to MHC molecules. The IP26 antibody is mitogenic and useful for the flow cytometric analysis of TCRαβ+ T-cell subsets.
The antibody was conjugated to BD Horizon™ Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon™ Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.
Development References (5)
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Agrawal SG, Marquet J, Plumas J, et al. Multiple co-stimulatory signals are required for triggering proliferation of T cells from human secondary lymphoid tissue. Int Immunol. 2001; 13(4):441-450. (Clone-specific: Activation, Functional assay, Stimulation). View Reference
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Nikolova M, Marie-Cardine A, Boumsell L, Bensussan A. BY55/CD160 acts as a co-receptor in TCR signal transduction of a human circulating cytotoxic effector T lymphocyte subset lacking CD28 expression. Int Immunol. 2002; 14(5):445-451. (Clone-specific: Flow cytometry). View Reference
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Oettgen HC, Kappler J, Tax WJ, Terhorst C. Characterization of the two heavy chains of the T3 complex on the surface of human T lymphocytes. J Biol Chem. 1984; 259(19):12039-12048. (Biology). View Reference
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Ortonne N, Huet D, Gaudez C, et al. Significance of circulating T-cell clones in Sezary syndrome. Blood. 2006; 107(10):4030-4038. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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van Dongen JJ, Lhermitte L, Böttcher S, et al. EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes. Leukemia. 2012; 26(9):1908-1975. (Clone-specific: Flow cytometry). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.