CD8 antibody

Catalog No. ABIN114175

Product Details anti-CD8 Antibody

Target: CD8

Reactivity: Mouse

Host: Rat

Clonality: Monoclonal

Conjugate: This CD8 antibody is un-conjugated

Application: Flow Cytometry (FACS), Immunohistochemistry (Frozen Sections) (IHC (fro))

Specificity: This anti-mouse CD8a antigen monoclonal antibody recognizes the mouse CD8α chain. The α chain of CD8 associates with the CD8β chain to form a CD8α/ β heterodimer that is expressed by the majority of thymocytes and by the MHC class I restricted subset of mature T cells1. Mouse CD8α can also form a CD8α/α chain homodimer on subsets of CD8 positive cells. For this reason antibodies specific for CD8a rather than CD8b are recommended for a rigorous delineation of CD8 positive cells.

Purification: Purified

Clone: CT-CD8a

Isotype: IgG2a

Application Details

Application Notes: Flow Cytometry Analysis (See Protocol). (Reported to be useful in immunohistochemistry on acetone fixed frozen sections. )
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation with Lympholyte®-M cell separation medium. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of thissuspension to each tube (each tube will then contain 1x10e6 cells, representing 1 test). 4. To each tube, add ~1. 0 µg* of this Ab. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4°C. 7. Wash 2 times at 4°C. 8. Add 100 µl of secondary antibody (FITC Goat anti-rat IgG (H+L)) at 1: 500 dilution. 9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes areprotected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C in media B. 11. Resuspend the cell pellet in 50 µl ice cold media B. 12. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidiumiodide at 0. 5 mg/ml in PBS. This stains dead cells by intercalating in DNA.

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

Buffer: PBS and 0.02 % Sodium Azide as preservative.

Preservative: Sodium azide

Precaution of Use: This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freezing and thawing.

Storage: 4 °C/-20 °C

Storage Comment: Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.

Target Details for CD8

Target: CD8

Alternative Name: CD8 (CD8 Products)

Background: The CD8 antigen is a cell surface glycoprotein found on most cytotoxic T lymphocytes that mediates efficient cell to cell interactions within the immune system. The CD8 antigen, acting as a coreceptor, and the T cell receptor on the T lymphocyte recognize antigen displayed by an antigen presenting cell (APC) in the context of class I MHC molecules. The functional coreceptor is either a homodimer composed of two alpha chains, or a heterodimer composed of one alpha and one beta chain. Both alpha and beta chains share significant homology to immunoglobulin variable light chains.Synonyms: CD8 alpha chain, CD8A, MAL, T-cell surface glycoprotein CD8 alpha chain, T-lymphocyte differentiation antigen T8/Leu-2

Gene ID: 12525

NCBI Accession: NP_001074579

UniProt: P01731