Description
The 53-7.3 monoclonal antibody specifically binds to a monomorphic epitope of CD5, a member of the scavenger receptor cysteine-rich protein superfamily and the major ligand of CD72, found on thymocytes, T lymphocytes, thymic NKT cells, and a subset of B lymphocytes, but not on NK cells or splenic NKT cells. The level of surface CD5 expression is developmentally regulated in the thymus, starting with low levels on CD4-CD8- thymocytes and increasing as they mature to CD4+CD8+ then CD4+CD8- or CD4-CD8+ thymocytes. Relatively high levels are maintained on peripheral T lymphocytes. The level of CD5 antigen detected on T helper cells has been reported to be somewhat higher than that on T cytotoxic/suppressor and B cells. Few, if any, intestinal intraepithelial lymphocytes bearing the γδ T-cell receptor express CD5. Phenotypic, anatomical, functional, developmental, and pathogenic characteristics of peripheral CD5+ B cells suggest that they may represent a distinct lineage, known as B-1 cells. The frequency of these CD5+ B cells has been reported to show strain-dependent variation. An additional population of CD5+ B lymphocytes resides in the thymus, where it matures from intrathymic B-cell progenitors. It has been proposed that CD5 is a costimulatory molecule which mediates interactions of cells in the immune system and negatively regulates signal transduction mediated by the T-cell receptor and B-cell receptor.
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. 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.
8. 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.
9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
10. An isotype control should be used at the same concentration as the antibody of interest.
11. 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.
