Purified Rat Anti- Mouse CD8a
Clone 53-6.7 (RUO)
- Brand BD Pharmingen™
- Alternative Name Cd8a; CD8 alpha chain; Ly-2; Lyt2; Lyt-2; Ly-35; Ly-B
- Concentration 62.5 µg/ml
- Isotype Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ
- Reactivity Mouse (QC Testing)
Flow cytometry (Routinely Tested)
Immunohistochemistry-frozen, Immunohistochemistry-zinc-fixed (Tested During Development)
Immunohistochemistry-formalin (antigen retrieval required) (Not Recommended)
- Immunogen Mouse Spleen Cells or Thymocyte Membranes
- Entrez Gene ID 12525
- Storage Buffer Aqueous buffered solution containing BSA, goat serum, and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The 53-6.7 monoclonal antibody specifically binds to the 38 kDa α and 34 kDa α' chains of the CD8 differentiation antigen (Ly-2 or Lyt-2) of all mouse strains tested. The CD8 α and α' chains (CD8a) form heterodimers with the CD8 β chain (CD8b, Ly-3, or Lyt-3) on the surface of most thymocytes. A subpopulation of mature T lymphocytes (i.e., MHC class I-restricted T cells, including most T suppressor/cytotoxic cells) expresses almost exclusively the CD8 αβ heterodimer. Subsets of γδ TCR-bearing T cells, intestinal intrapithelial lymphocytes, and dendritic cells express CD8a without CD8b. It has been suggested that the expression of the CD8a/CD8b heterodimer is restricted to T lymphocytes which matured in the thymus or in an extrathymic environment that had been influenced by thymus-initiated neuroendocrine signals. CD8 is an antigen coreceptor on the T-cell surface which interacts with MHC class I molecules on antigen-presenting cells or epithelial cells. It participates in T-cell activation through its association with the T-cell receptor complex and protein tyrosine kinase lck (p56 [lck]). The CD8 α and α' chains arise from alternatively spliced messengers of a single CD8a gene. The longer α form associates with p56 [lck] via a CXCP motif in its cytoplasmic domain, which it shares with CD4, but not with CD8b. The truncated α' chain is unable to associate with p56 [lck], and it may function to attenuate the CD8-mediated costimulatory signal during intrathymic T-cell maturation. In vivo and in vitro treatment with 53-6.7 mAb has reportedly been effective at depleting CD8+ peripheral T lymphocytes. The 53-6.7 antibody has also been reported to cross-react with CD8 α- and α'-like polypeptides on subsets of thymic and peripheral lymphocytes in the Egyptian toad, Bufo regularis.
- Format Purified
Suggested Companion Products
Preparation and Storage
Store undiluted at 4°C.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- This antibody has been developed for the immunohistochemistry application. However, a routine immunohistochemistry test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
Immunohistochemistry: Clone 53-6.7 is recommended to test for immunohistochemical staining of acetone-fixed frozen sections or zinc-fixed paraffin sections of mouse spleen or thymus. IHC of formalin-fixed paraffin embedded sections is not recommended. The antibody has been reported to stain the membranes of thymocytes and a subpopulation of mature T lymphocytes that are MHC class I restricted. The isotype control recommended for use with this antibody is purified rat IgG2a (Cat. No. 559073). For optimal indirect immunohistochemical staining, the 53-6.7 antibody should be titrated (1:10 to 1:50 dilution and visualized via a three-step procedure in combination with Biotin Goat Anti-Rat Ig (Cat. No. 559286) as the secondary antibody and Streptavidin-HRP (Cat. No. 550946) together with the DAB detection system (Cat. No. 550880). More conveniently, the anti-rat Ig HRP detection kit (Cat. No. 551013) can be used which contains the biotinylated secondary antibody, the antibody diluent, streptavidin-HRP and a DAB substrate for use in the staining procedure.