Cd8a Mouse Monoclonal Antibody [Clone ID: AD4(15)]

Specifications

Product Data
Clone Name	AD4(15)
Applications	CT, FC
Recommend Dilution	Cytotoxicity assays. Flow Cytometry. Results: Tissue Distribution by Flow Cytometry Analysis: Mouse Strain: BALB/c Cell Concentration : 1x106 cells per test. Antibody Concentration Used: 2.0 μg /106 cells. Isotypic Control: Mouse IgM. Cell Source (Percentage of cells stained above control): Spleen (10.2%), Thymus (66.0%).
Reactivity	Mouse
Host	Mouse
Clonality	Monoclonal
Immunogen	C57BL/6 Donor: B6-Ly-2a spleen Fusion Partner: Myeloma P3/X63-Ag8
Specificity	Anti-mouse CD8a (Ly 2.2) monoclonal antibody reacts with a sub-population of T-lymphocytes from Mouse strains expressing the Ly-2.2 phenotype but does not react with lymphocytes from strains expressing the Ly-2.1 phenotype.
Formulation	PBS State: Purified State: Liquid purified IgG fraction Preservative: 0.02% Soidium Azide
Concentration	lot specific
Purification	Euglobin precipitation
Conjugation	Unconjugated
Storage Condition	Store 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 2x107 cells/ml in media A. Add 50 μl of this suspension to each tube (each tube will then contain 1x106 cells, representing 1 test). 4. To each tube, add 2.0 μg of this antibody Cat.-No CL010P. 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-mouse IgM-3 (H+L)) at 1/500 dilution. 9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes are protected 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 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).
Reference Data

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