CD8 antibody (Biotin)

Catalog No. ABIN114164

The Rat Monoclonal anti-CD8 antibody has been validated for FACS. It is suitable to detect CD8 in samples from Mouse.

Quick Overview for CD8 antibody (Biotin) (ABIN114164)

Target: CD8

Reactivity: Mouse

Host: Rat

Clonality: Monoclonal

Conjugate: This CD8 antibody is conjugated to Biotin

Application: Flow Cytometry (FACS)

Clone: YTS169-4

Product Details anti-CD8 Antibody (Biotin)

Specificity: This anti-mouse CD8a (Ly 2) monoclonal antibody reacts with a protein of approximately 30 kDa found on mouse thymocytes and mouse cytotoxic/ suppressor T cells. It does not bind to mouse helper/inducer T cells. It binds to T lymphocytes from all mouse strains regardless of phenotypic expression (i.e. reacts with T lymphocytes from mouse strains expressing the Ly 2.1 or Ly 2.2 phenotype). It can be used to investigate the role of T cells in models for infectious disease, autoimmunity, transplantation tolerance and fundamental aspects of immunology.

Purification: Protein G Chromatography

Immunogen: Murine thymocytes Donor: (LOU X DA) F1 rats Fusion Partner: Myeloma Y3/Ag 1.2.3

Isotype: IgG2b

Application Details

Application Notes: Flow Cytometry Analysis (see Protocols).
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 1 x 10e6 cells, representing 1 test). 4. To each tube, add 0. 1-0. 2 µg* of this Ab per 10e6 cells. 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 (Streptavidin-FITC) at a 1: 500 dilution. 9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protectedfrom light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C. 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. The stains dead cells by intercalating in DNA. Media: A. Phosphate buffered saline (pH 7. 2) + 5% normal serum of host species + sodium azide(100 µl of 2M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7. 2) + 0. 5% Bovine serum albumin + sodium azide (100µl of 2M sodium azide in 100 mls). Results - Tissue DistributionMouse Strain: BALB/cCell Concentration: 1x10e6 cells per testsAntibody Concentration Used: 0. 2 µg/10e6 cellsIsotypic Control: Biotin Rat IgG2bResults - Strain DistributionCell Concentration: 1x10e6 cells per testsAntibody Concentration Used: 0. 2 µg/10e6 cellsStrains Tested: BALB/c, CBA/J, C3H/He,C57BL/6Positive: BALB/c, CBA/J, C3H/He,C57BL/6Negative: none

Restrictions: For Research Use only

Handling

Concentration: 0.1 mg/mL

Buffer: PBS, 0.02 % NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/mL

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

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