MHC Class II I-Abd Mouse Monoclonal Antibody [Clone ID: 28-16-8S]

Specifications

Product Data
Clone Name	28-16-8S
Applications	FC
Recommend Dilution	Flow Cytometry (See Protocols).
Reactivity	Mouse
Host	Mouse
Clonality	Monoclonal
Immunogen	C3H.SW spleen Donor: C3H Fusion Partner: SP2/0Ag.14
Specificity	Recognizes Mouse MHC class II I-Ab and cross reacts with I-Ad. This Monoclonal Antibody reacts with the I-Ab encoded MHC class II antigen expressed on Mouse strains of the H-2b haplotype. It also reacts with the I-Ad encoded MHC class II antigen expressed on Mouse strains of the H-2d haplotype. Class II antigens are most highly expressed on antigen presenting cells including B cells, macrophages, dendritic cells and certain epithelial cells.
Formulation	PBS containing 0.02% Sodium Azide as a preservative as a and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml Label: PE State: Liquid purified IgM 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. Avoid repeated freezing and thawing.
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 1x10e6 cells, representing 1 test). 4. To each tube, add~0.2 µg of CL067R 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 ml of 2M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7.2) + 0.5% Bovine serum albumin + sodium azide (100 ml of 2M sodium azide in 100 mls). Results: Tissue Distribution by Flow Cytometry Analysis: Mouse strain: C57BL/6 Cell Concentration: 1x10e6 cells per test. Antibody Concentration Used: 0.2 µg/10e6 cells. Isotypic Control: PE Mouse IgM Cell Source: Percentage Stained Above Control: Thymus: 28.2% Spleen: 40.7%
Reference Data

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