Description
The J43 monoclonal antibody specifically recognizes CD279 which is also known as PD-1 (programmed death-1). CD279 is a 50-55-kDa glycoprotein encoded by the Pdcd1 gene of the CD28 family of the Ig superfamily. The expression of Pdcd1 mRNA and PD-1 protein is tightly regulated. PD-1 is transiently expressed on CD4-CD8 thymocytes, it is upregulated on some cell lines upon induction of apoptosis, it is induced on thymocytes and splenic T and B lymphocytes after stimulation through their antigen receptors, and it is induced on activated myeloid cells. In addition, Pdcd1 mRNA is transiently expressed in developing B lymphocytes at the pro-B-cell stage. The presence of an ITIM (Immunoreceptor Tyrosine-based Inhibitory Motif) on PD-1's intracytoplasmic region and the development of splenomegaly and breakdown of peripheral tolerance in PD-1[-/-] mice suggest that PD-1 is involved in the negative regulation of immune responses. The PD-1 ligands, B7-H1 (also known as PD-L1, CD274) and B7-DC (PD-L2, CD273), are members of the B7 family of the Ig superfamily. The J43 antibody blocks the binding of PD-1 to its two ligands.
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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
4. An isotype control should be used at the same concentration as the antibody of interest.
5. Researchers should determine the optimal concentration of this reagent for their individual applications.
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. 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.
8. 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.
9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
10. 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.
11. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
12. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
