Description
The L128 monoclonal antibody specifically binds to human CD27. CD27 is a 55-kDa disulfide-linked dimer that is a member of the nerve growth factor (NGF) super family. This family also includes CD40, rat OX40, tumor necrosis factor (TNF) receptors and CD95 (Fas). With its ligand CD70, CD27 acts in a co-stimulatory fashion on T lymphocytes. Present on most peripheral blood T lymphocytes and medullary thymocytes, the CD27 antigen is upregulated upon activation with the release of a soluble form, 28 to 32 kDa. It is also detected on a subpopulation of approximately 33% of circulating B lymphocytes. Following exposure to antigens, CD45RA+ T lymphocytes respond by upregulating the CD27 antigen. After maximal stimulation, the CD27 antigen cannot be re-expressed on long-term cultures or on CD45RA-CD27+ T lymphocytes. The CD4+CD27- population is contained within the memory CD45RO+ subset that proliferates after exposure to allergens. Two subpopulations of B lymphocytes bearing the CD27 antigen secrete IgM (δ+) and IgG (δ-).
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.
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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. 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.
3. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
5. An isotype control should be used at the same concentration as the antibody of interest.
6. CF™ is a trademark of Biotium, Inc.
7. 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.
8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
9. 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.
10. For U.S. patents that may apply, see bd.com/patents.
11. 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.
12. 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.
13. 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.
