MHC Class II RT1D Mouse Monoclonal Antibody [Clone ID: OX-17]

Specifications

Product Data
Clone Name	OX-17
Applications	FC
Recommend Dilution	Flow Cytometry.
Reactivity	Rat
Host	Mouse
Clonality	Monoclonal
Immunogen	Rat spleen membrane glycoproteins depleted of Ia-A antigens. Immunocyte Donor: BALB/c spleen Fusion Partner: X63 Ag8.653
Specificity	This monoclonal antibody recognizes a monomorphic determinant on the α chain of the rat Ia antigen and appears to be the rat homologue of mouse Ia-E. It recognizes the rat Ia product present on B, but not T cells from lymph node or thoracic duct lymph. It does not bind to thymocytes or erythrocytes. The antibody does not cross-react with rat Ia-A or mouse Ia-E antigen, but rabbit antibody raised against the antibody affinity column-purified MRC OX-17 antigen cross-reacted on tissues of mice expressing Ia-E mouse antigen but not on those mouse strains that were Ia-E antigen negative.
Formulation	PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml Label: FITC State: Liquid purified Ig
Concentration	lot specific
Purification	Protein G Chromatography
Conjugation	FITC
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. This antibody is photosensitive and should protected from light.
Synonyms	HLA Class II
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®-Rat 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 0.05-0.1 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. (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: Rat Strain: Fischer Cell Concentration: 1x10e6 cells per test Antibody Concentration Used: 0.1 µg/10e6 cells Isotypic Control: FITC Mouse IgG1 Cell Source Percentage of cells stained above control: Thymus 10.2% Spleen 49.0% Lymph Node 27.9%
Reference Data

*Delivery time may vary from web posted schedule. Occasional delays may occur due to unforeseen complexities in the preparation of your product. International customers may expect an additional 1-2 weeks in shipping.