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Description: The 53-6.7 monoclonal antibody reacts with the mouse CD8a molecule. CD8a is 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 alpha beta TCR T cells express CD8 alpha beta while gamma delta TCR T cells, a subpopulation of intestinal intraepithelial lymphocytes (IELs) and dendritic cells express CD8 alpha alpha. 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.
Applications Reported: This 53-6.7 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This 53-6.7 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.5 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Super Bright 780 can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter, or equivalent. Please make sure that your instrument is capable of detecting this fluorochrome.
When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.
In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).
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) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) 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: 405 nm; Emission: 780 nm; Laser: Violet Laser
Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
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.
Protein Aliases: CD8a; CD8alpha; CD8b; CD8beta; fCD8; Leu-2; leu-2a; Lymphocyte antigen 3; Lyt-2.1 lymphocyte differentiation antigen (AA at 100); T-cell membrane glycoprotein Ly-3; T-cell surface glycoprotein CD8 alpha chain; T-cell surface glycoprotein CD8 beta chain; T-cell surface glycoprotein Lyt-2; T-cell surface glycoprotein Lyt-3
Gene Aliases: BB154331; Cd8a; Cd8b; Cd8b1; Ly-2; Ly-3; Ly-35; Ly-B; Ly-C; Lyt-2; Lyt-3; Lyt2; Lyt3