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Two-color flow cytometric analysis of TCR Vβ5 Related expression on Human peripheral blood T lymphocytes. Human peripheral blood cells were stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426) and with either Alexa Fluor™ 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565357; Left Plot) or Alexa Fluor™ 647 Mouse Anti-Human TCR Vβ5 Related antibody (Cat. No. 568627/568628; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing the correlated expression of TCR Vβ5 Related (or Ig Isotype control staining) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of viable human lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
BD Pharmingen™ Alexa Fluor™ 647 Mouse Anti-Human TCR Vβ5 Related
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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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
The MEM-262 monoclonal antibody recognizes an epitope on the extracellular region of the beta chain of the human αβ T cell receptor for antigen (TCR) expressed by a TCR Vβ5.3+ human HPB-ALL cell line as well as a small subset of peripheral blood T cells. The peripheral T cell subset recognized by the MEM-262 antibody is larger than that recognized by other TCR Vβ5.3-specific antibodies and so the recognized epitope may be referred to as TCR Vβ5 Related. This antibody is useful for multiparameter analyses designed to study the nature of MEM-262-positive T cells including normal T cells and T cell clones or T cell lines. It is also useful for analyzing TCR Vβ repertoires expressed by T cell populations collected from blood, tissues or other sources in health and disease models. This monoclonal antibody can reportedly stimulate the proliferation of MEM-262-positive T cells.
Development References (3)
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Gertow J, Berglund S, Okas M, et al. Characterization of long-term mixed donor-donor chimerism after double cord blood transplantation.. Clin Exp Immunol. 2010; 162(1):146-55. (Clone-specific: Flow cytometry). View Reference
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Giudice V, D'Addona M, Montuori N, Selleri C. The Value of Flow Cytometry Clonality in Large Granular Lymphocyte Leukemia.. Cancers (Basel). 2021; 13(18):4513. (Clone-specific: Flow cytometry). View Reference
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Pavlistová D, Drbal K, Hilgert I, Horejsí V. A novel monoclonal reagent recognizing native and denatured Vbeta5.3-related chains of human T cell receptor.. Immunol Lett. 2003; 88(2):105-8. (Immunogen: Flow cytometry, Functional assay, Immunoaffinity chromatography, Immunoprecipitation, Stimulation). View Reference
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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.