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Description: The OKT8 monoclonal antibody reacts with the human CD8a molecule, an approximately 32-34 kDa cell surface receptor expressed either as a heterodimer with the CD8 beta chain (CD8 alpha beta) or as a homodimer (CD8 alpha alpha). A majority of thymocytes and a subpopulation of mature T cells and NK cells express CD8a. CD8 binds to MHC class I and through its association with protein tyrosine kinase p56lck plays a role in T-cell development and activation of mature T cells. Preliminary testing indicates that OKT8 and two other mouse anti-human CD8 antibodies (clone RPA-T8, Product # 14-0088 and clone HIT8a, Product # 14-0089-82) do not compete with each other for binding to human peripheral blood leukocytes by flow cytometric analysis, suggesting that they do not bind to similar epitopes or block each other by steric hindrance.
Applications Reported: This OKT8 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This OKT8 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
PerCP-eFluor 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.
Cluster of differentiation 8 (CD8), a type I transmembrane glycoprotein of the immunoglobulin family of receptors, plays an integral role in signal transduction, and T cell differentiation and activation. CD8 is predominantly expressed on T cells as a disulfide-linked heterodimer of CD8alpha and CD8beta, where it functions as a co-receptor, along with T cell receptor (TCR), for major histocompatibilty complex class I (MHC-I) molecules; whereas its counterpart, CD4, acts as a co-receptor for MHC-II molecules. CD8 exists on the cell surface, where the CD8alpha chain is essential for binding to MHC-I. CD8 is also expressed on a subset of T cells, NK cells, monocytes and dendritic cells as disulfide-linked homodimers of CD8alpha. Ligation of MHC-I/peptide complexes presented by antigen-presenting cells (APCs), triggers the recruitment of lymphocyte-specific protein tyrosine kinase (Lck), which leads to lymphokine production, motility and cytotoxic T lymphocyte (CTL) activation. Once activated, CTLs play a crucial role in the clearance of pathogens and tumor cells. Differentiation of naive CD8+ T cells into CTLs is strongly enhanced by IL-2, IL-12 and TGF-beta1.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Protein Aliases: CD8 antigen, alpha polypeptide (p32); CD8 antigen, beta polypeptide 1 (p37); CD8a; CD8alpha; CD8b; CD8beta; fCD8; Leu-2; leu-2a; Leu2 T-lymphocyte antigen; OKT8 T-cell antigen; T cell co-receptor; T lymphocyte surface glycoprotein beta chain; T-cell antigen Leu2; T-cell surface glycoprotein CD8 alpha chain; T-cell surface glycoprotein CD8 beta chain; T-lymphocyte differentiation antigen T8/Leu-2; T8 T-cell antigen
Gene Aliases: CD8; CD8A; CD8B; CD8B1; LEU2; LY3; LYT3; MAL; p32; P37
UniProt ID: (Human) P01732, (Human) P10966
Entrez Gene ID: (Human) 925, (Human) 926
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