Description
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.
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.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.
1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
3. For U.S. patents that may apply, see bd.com/patents.
4. 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.
5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
6. 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.
7. 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.
8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
9. An isotype control should be used at the same concentration as the antibody of interest.
10. 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.
11. 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.
