Cd8a Rat Monoclonal Antibody [Clone ID: YTS169.4]

Specifications

Product Data
Clone Name	YTS169.4
Applications	FC
Recommend Dilution	Flow Cytometry Analysis (see Protocols). This antibody has been reported to be suitable for use in Immunohistochemistry on PFA-Fixed Paraffin-Embedded tissue sections (antigen retrieval is required). (7) This clone has been reported to work in Immunohistochemistry on Frozen Sections. (6)
Reactivity	Mouse
Host	Rat
Clonality	Monoclonal
Immunogen	Mouse Ly-2 thymocytes. Donor: (LOU X DA) F1 rat. Fusion Partner: Y3/Ag1.2.3.
Specificity	This anti-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 (ie. 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. (1) It can also be useful to identify/eliminate cytotoxic or suppressor T lymphocytes in vivo or in vitro. Strains Tested: C57BL/6, BALB/c, AKR/J, C3H/He Positive: : C57BL/6, BALB/c, AKR/J, C3H/He Negative: None.
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: PE State: Liquid purified IgG fraction
Concentration	lot specific
Conjugation	PE
Storage Condition	Store the antibody undiluted at 2-8°C. DO NOT FREEZE! This product is photosensitive and should be protected from light.
Gene Name	CD8 antigen, alpha chain
Database Link	Entrez Gene 12525 Mouse UniProt ID P01731
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
Note	Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare cell suspension in Media A. For cell preparations, deplete the red blood cell population. 2. Wash 2 times. 3. Resuspend cells to a concentration of 2x10e7 cells/ml Media A. Add 50 µl of this suspension to each tube (each tube will then contain 1x10e6 cells, representing 1 test. 4. To each tube add 2.0 µg of this Ab per 10e6 cells. 5. Vortex to ensure through mixing of antibody and cells. 6. Incubate tubes at 4°C for 30 minutes. (It is recommended that the tubes are protected from light, since most fluorochromes are light sensitive.) 7. Wash 2 times at 4°C in Media B. 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 phosphate buffered saline. (This stains dead cells by intercalating DNA). Media: A. Phosphate buffered saline (pH 7.2) + 5% normal serum of host species + sodium azide (100 ul of 2 M sodium azide in 100 mls.) B. Phosphate buffered saline (pH 7.2) + 0.5% bovine serum albumin + sodium azide (100 ul of 2 M sodium azide in 100 mls.) Results - Tissue Distribution Mouse Strain: BALB/c Cell Concentration: 1x10e6 cells per test Antibody Concentration Used: 2.0 µg/10e6 cells Isotypic Control: PE Rat IgG2b,k Percentage of cells stained above control: Thymus 66.2% Spleen 19.6 % Lymph node 14.0% Results - Strain Distribution Cell Concentration: 1x10e6 cells per test Antibody Concentration Used: 0.2 µg/10e6 cells Strains Tested: BALB/c, C57BL/6 Positive: BALB/c, C57BL/6 Negative: none
Reference Data

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