Cd8a Mouse Monoclonal Antibody [Clone ID: 49-31.1]

Specifications

Product Data
Clone Name	49-31.1
Applications	FC
Recommend Dilution	Flow Cytometry.
Reactivity	Mouse
Host	Mouse
Clonality	Monoclonal
Immunogen	Immunization: Recipient: 129/ReJ Donor: CBA Fusion Partner: Spleen from immunized recipient fused with Myeloma P3 NSI-Ag 4-1
Specificity	Anti-mouse Ly-2.1 monoclonal antibody reacts with a sub-population of lymphocytes from mouse strains expressing the Ly 2.1 (CD8a) phenotype, but does not react with lymphocytes from mouse strains expressing the Ly 2.2 phenotype.
Formulation	PBS containing 0.02% sodium azide (NaN3) as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml. Label: Biotin State: Liquid purified Ig fraction
Concentration	lot specific
Purification	Affinity chromatography on Protein G
Conjugation	Biotin
Storage Condition	Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing.
Gene Name	CD8 antigen, alpha chain
Database Link	Entrez Gene 12525 Mouse UniProt ID P01731
Synonyms	CD8 alpha chain, CD8A, MAL
Note	Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population 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 this suspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube, add 0.5-0.1 μg of this antibody 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-PE) at a 1/500 dilution. 9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protected from 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 propidium iodide at 0.5 mg/ml in PBS. This 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 (100 μl of 2M sodium azide in 100 mls). Results: Tissue Distribution by Flow Cytometry Analysis: Mouse Strain: C3H/He Cell Concentration: 1x10e6 cells per test Antibody Concentration Used: 0.2 μg/10e6 cells Percentage of cells stained above control: Thymus 80.7%
Reference Data

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