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

Specifications

Product Data
Clone Name	AD4(15)
Applications	FC
Recommend Dilution	Cytotoxicity assays. Flow Cytometry.
Reactivity	Mouse
Host	Mouse
Clonality	Monoclonal
Immunogen	C57BL/6 Donor: B6-Ly-2a Fusion Partner: Myeloma line P3/X63Ag8
Specificity	Anti-mouse CD8a (Ly 2.2) monoclonal antibody reacts with a subpopulation 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.
Isotype	IgM
Formulation	PBS containing 0.02% Sodium Azide and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml. Label: FITC State: Liquid purified Ig fraction Label: Fluorescein isothiocyanate isomer 1
Purification	Euglobin precipitation
Conjugation	FITC
Storage Condition	Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. This product is photosensitive and should be protected from light. 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 2.0-1.0 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. (It is recommended that the tubes are protected from light, since most fluorochromes are light sensitive.) 7. Wash 2 times at 4°C. 8. Resuspend the cell pellet in 50 μl ice cold media B. 9. 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: BALB/c Cell Concentration : 1x10e6 cells per tests Antibody Concentration Used: 2.0 μg/10e6 cells Isotypic Control: FITC Mouse IgM Percentage of cells stained above control: Spleen 10.2% Thymus 66.0%
Reference Data

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