MHC Class II I-Ab Mouse Monoclonal Antibody [Clone ID: 25-5-16S]

Specifications

Product Data
Clone Name	25-5-16S
Applications	FC
Recommend Dilution	Flow Cytometry (See Protocols).
Reactivity	Mouse
Host	Mouse
Clonality	Monoclonal
Specificity	Recognizes Mouse MHC class II I-Ab, private for I-Ab. This Monoclonal antibody specifically reacts with the I-Ab encoded MHC class II antigen expressed on Mouse strains of the H-2b haplotype.
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: FITC State: Liquid purified IgM fraction
Concentration	lot specific
Conjugation	FITC
Storage Condition	Store the antibody 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.
Database Link	UniProt ID P18468
Background	Class II antigens are most highly expressed on antigen-presenting cells including B cells, macrophages, dendritic cells and certain epithelial cells.
Synonyms	H2-Eb1, H-2 class II histocompatibility antigen I-A beta chain
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~1.0 µg of SM080FS 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: 1.0 µg/10e6 cells. Isotypic Control: FITC Mouse IgM
Reference Data

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