MHC Class I H-2 Kb / H-2 Db Mouse Monoclonal Antibody [Clone ID: 5041.16.1]

Specifications

Product Data
Clone Name	5041.16.1
Applications	FC
Recommend Dilution	Flow cytometry (see protocol below).
Reactivity	Mouse
Host	Mouse
Clonality	Monoclonal
Specificity	This antibody is specific for cells expressing the H-2K antigen coded for by the b haplotype and for cells expressing the H-2D antigen coded for by the b haplotype.
Formulation	PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml. Label: PE State: Liquid Ig fraction
Concentration	lot specific
Purification	Protein G chromatography
Conjugation	PE
Storage Condition	Store at 2 - 8 °C. DO NOT FREEZE. This product is photosensitive and should be protected from light.
Synonyms	HLA Class I
Note	Protocol: FLOW CYTOMETRY ANALYSIS: 1. Prepare cell suspension in Media A. For cell preparations, deplete the red blood cell population. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50µl of this suspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube add 0.2- 0.5 mg of 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. 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 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 µl of 2 M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7.2) + 0.5% bovine serum albumin + sodium azide (100 µl of 2 M sodium azide in 100 mls). FLOW CYTOMETRIC ANALYSIS: Donor: C57BL/6 Cell Concentration: 1x10e6 cells Antibody Concentration: 0.2 µg/10e6 cells Isotypic Control: PE Mouse IgG2a, k (see picture below) STRAIN DISTRIBUTION: Procedure: As above Antibody Concentration: 0.2µg/10e6 cells Strains Tested: Strain Haplotype +/- BALB/c H-2d - C3H/He H-2k - CBA/J H-2k - C57BL/6 H-2b + B6Lyt 2.1 3.1 H-2b +
Reference Data

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