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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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
Companion Products
The MH3-2 monoclonal antibody specifically recognizes the human variable beta 5.2 (TCR Vβ5.2) and variable beta 5.3 (TCR Vβ5.3) domains of the beta subunit for the αβ T cell receptor (TCR Vβ5.2/5.3). The TCR Vβ5.2 and TCR Vβ5.3 domains are encoded by the TRBV5-6 (T cell receptor beta variable 5-6; also known as TCRBV5S2) and TRBV5-5 (T cell receptor beta variable 5-5; TCRBV5S3) gene segments of the multimembered Vβ5 subfamily within the TRB (T cell receptor beta locus), respectively. Functional T-cell receptor beta chains (TCR-β) are generated by genomic rearrangement of TCR variable (Vβ), diversity (Dβ), joining (Jβ) and constant (Cβ) region gene segments by precursor T cells. Distinct αβ TCR containing either TCR Vβ5.2 or TCR Vβ5.3 are clonally expressed on subsets of thymocytes or peripheral CD4+ or CD8+ T cells. These TCR β beta chains can heterodimerize with TCR α chains to form αβ TCR that function as signaling T cell receptors for antigen. Human αβ TCR recognize peptides presented by HLA (MHC) molecules and in conjunction with the associated CD3 complex, can transduce intracellular signals that initiate precursor or mature T cell responses. The MH3-2 antibody may be useful for analyzing the frequencies or numbers of TCR Vβ5.2/5.3-positive thymocytes or mature T cells as well as the levels of TCR Vβ5.2/5.3 expressed by these cells. The MH3-2 antibody can be used in research applications such as flow cytometry to help characterize the TCR Vβ repertoires of precursor or mature T cell populations.
Development References (7)
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Folch G, Jabado-Michaloud J, Lefranc M-P. Reagents monoclonal antibodies: anti-human TRBV, IMGT repertoire, IMGT Web resources. http://www.imgt.org/IMGTrepertoire/Regulation/antibodies/human/TRB/TRBV/Hu_TRBVMab.html. IMGT®. 2020; 4. (Clone-specific: Flow cytometry).
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Kabelitz D, Wesch, D. T-Cell Receptor Analysis by Flow Cytometry. In: Sack U, Tarnok A, Rothe G. Sack U, Tarnok A, Rothe G, ed. Basic Principles, Methods and Clinical Applications of Flow Cytometry. New York: Karger, Basel; 2009:200-210. View Reference
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Labrecque N, McGrath H, Subramanyam M, Huber BT, Sékaly RP. Human T cells respond to mouse mammary tumor virus-encoded superantigen: V beta restriction and conserved evolutionary features.. J Exp Med. 1993; 177(6):1735-43. (Clone-specific: Flow cytometry). View Reference
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Oh S, Terabe M, Pendleton CD, et al. Human CTLs to wild-type and enhanced epitopes of a novel prostate and breast tumor-associated protein, TARP, lyse human breast cancer cells.. Cancer Res. 2004; 64(7):2610-8. (Clone-specific: Flow cytometry). View Reference
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Posnett DN, Romagne F, Necker A, Kotzin BL, Sékaly R-P. First human TCR monoclonal antibody workshop. The Immunologist. 1996; 4(1):5-8. (Clone-specific).
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Soudeyns H, Champagne P, Holloway CL, et al. Transient T cell receptor beta-chain variable region-specific expansions of CD4+ and CD8+ T cells during the early phase of pediatric human immunodeficiency virus infection: characterization of expanded cell populations by T cell receptor phenotyping.. J Infect Dis. 2000; 181(1):107-20. (Clone-specific: Flow cytometry). View Reference
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Vonderheid EC, Boselli CM, Conroy M, et al. Evidence for restricted Vbeta usage in the leukemic phase of cutaneous T cell lymphoma.. J Invest Dermatol. 2005; 124(3):651-61. (Clone-specific: Flow cytometry). View Reference
Please refer to Support Documents for Quality Certificates
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.