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.
- 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.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- 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.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- 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.
Companion Products
The C21 monoclonal antibody specifically recognizes the variable beta 11 region of the β subunit of the human αβ T cell receptor for antigen (TCR Vβ11). TCR Vβ11 is encoded by TRBV25-1 (T cell receptor beta variable 25-1) which is also known as TCRBV11S1. TCR Vβ11 can pair with different TCR Vα region containing TCR α chains. The paired expression of semivariant TCR Vβ11 with invariant TCR Vα24 and Jα18 identifies some CD3+TCR αβ+ positive thymocytes and from ~ 0.01-0.92% of peripheral blood CD3+ TCR αβ+ T cells known as invariant NKT (iNKT) cells or Type I NKT cells. iNKT cells can be divided into three subpopulations which are either CD4+, CD8+, or CD4-CD8- double negative. Due to their innate-like responsiveness, tissue resident iNKT cells are capable of rapid effector responses to microbial or self-lipid antigens presented by the cell surface MHC class Ib molecule, CD1d. iNKT cells recognize and respond to α-galactosylceramide (α-GalCer) and various self-lipids such as sphingolipids and diacylglycerols and microbial lipids presented by CD1d. iNKT cells can produce high levels of cytokines, including IL-4 and IFN-γ, and act as cytotoxic T cells against CD1d+ target cells following stimulation. The C21 antibody is useful for multiparameter cell sorting and phenotypic analyses designed to study the nature of TCR Vβ11-positive T cells and T cell clones. When combined with other TCR V region-specific antibodies, C15 is helpful for characterizing TCR Vβ repertoires expressed by T cell populations collected from blood, tissues or other sources in health and disease models including inflammation, infectious diseases, autoimmunity, and tumors.
Development References (6)
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Cossarizza A, Chang HD, Radbruch A, et al. Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition).. Eur J Immunol. 2021; 51(12):2708-3145. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Dellabona P, Padovan E, Casorati G, Brockhaus M, Lanzavecchia A. An invariant V alpha 24-J alpha Q/V beta 11 T cell receptor is expressed in all individuals by clonally expanded CD4-8- T cells.. J Exp Med. 1994; 180(3):1171-6. (Immunogen: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Kojo S, Adachi Y, Keino H, Taniguchi M, Sumida T. Dysfunction of T cell receptor AV24AJ18+, BV11+ double-negative regulatory natural killer T cells in autoimmune diseases.. Arthritis Rheum. 2001; 44(5):1127-38. (Clone-specific: Flow cytometry). View Reference
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Lal KG, Leeansyah E, Sandberg JK, Eller MA. OMIP-046: Characterization of invariant T cell subset activation in humans.. Cytometry A. 2018; 93(5):499-503. (Clone-specific: Flow cytometry). View Reference
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Oishi Y, Sakamoto A, Kurasawa K, et al. CD4-CD8- T cells bearing invariant Valpha24JalphaQ TCR alpha-chain are decreased in patients with atopic diseases.. Clin Exp Immunol. 2000; 119(3):404-11. (Clone-specific: Flow cytometry). View Reference
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Thomas SY, Hou R, Boyson JE, et al. CD1d-restricted NKT cells express a chemokine receptor profile indicative of Th1-type inflammatory homing cells. J Immunol. 2003; 171(5):2571-2580. (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.