Description: The HIT8a 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.
Applications Reported: This Hit8a antibody has been reported for use in flow cytometric analysis.
Applications Tested: This Hit8a 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.
Super Bright 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.
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).
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.
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: 405 nm; Emission: 780 nm; Laser: Violet Laser
Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
CD8 molecule is composed of two chains termed alpha and beta. CD8 is found on a T cell subset of normal cytotoxic / suppressor cells which make up approximately 20 to 35% of human peripheral blood lymphocytes. The CD8 antigen is also detected on natural killer cells, 80% of thymocytes, on a subpopulation of 30% of peripheral blood null cells and 15 to 30% of bone marrow cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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