Description
The YTH 76.3rMAb is a recombinant monoclonal antibody that specifically recognizes a monomorphic epitope on the extracellular region of the Human Leukocyte Antigen-B (HLA-B) heavy chain. This major histocompatibility complex (MHC) class I antigen is comprised of a polymorphic HLA-B heavy chain, a ~45 kDa type I transmembrane glycoprotein encoded by HLA-B, that is noncovalently associated with the invariant ~12 kDa Beta-2 (β2)-microglobulin light chain encoded by B2M. Hundreds of HLA-B alleles have been reported. HLA-B is expressed on most nucleated cells and leucocytes including specialized antigen presenting cells such as thymic epithelial cells and dendritic cells. HLA-B plays a major role in the MHC-restricted presentation of small peptides, including those derived from self or foreign antigens, that can be bound by TCR expressed on CD8+ thymocytes or T cells leading to either immune tolerance, the maturation of naïve CD8+ T cells, or the activation and differentiation of CD8+ cytotoxic effector T cells or memory CD8+ T cells. HLA-B also functions as a ligand for regulatory CD8 coreceptor molecules and some CD158 molecules that serve as regulatory MHC class I antigen receptors expressed by CD8+ T cells and NK cells. The YTH 76.3rMAb is derived from the hybridoma clone YTH 76.3, with Rat IgG2a, κ isotype. YTH 76.3rMAb has variable region sequences from the original YTH 76.3 clone appended to Rat IgG1 and kappa constant region sequences.
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. 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.
2. Researchers should determine the optimal concentration of this reagent for their individual applications.
3. 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.
4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
6. An isotype control should be used at the same concentration as the antibody of interest.
7. 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.
8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
10. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
12. 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.
