MHC Class I H2 Kd/Dd Mouse Monoclonal Antibody [Clone ID: 34-1-2S]

Specifications

Product Data
Clone Name	34-1-2S
Applications	FC
Recommend Dilution	Flow Cytometry.
Reactivity	Mouse
Host	Mouse
Clonality	Monoclonal
Immunogen	BDF splenocytes Donor: C3H spleen Fusion Partner: Sp2/0-Ag14
Specificity	Anti H-2KdDd mAb reacts with both H-2Kd and H-2Dd products. The antibody also cross reacts with Kbsrpq. This K-D cross reaction indicates the presence of shared specificities between the two separate H-2 regions.
Formulation	PBS containing 0.02% sodium azide (NaN3) as preservative and EIA grade BSA as a stabilizing protein Label: PE State: Liquid purified Ig fraction Label: R - Phycoerythrin
Concentration	lot specific
Purification	Affinity chromatography on Protein G
Conjugation	PE
Storage Condition	Store the antibody undiluted at 2-8°C. DO NOT FREEZE!
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 1 x 10e6 cells, representing 1 test). 4. To each tube, add 0.2-0.5 μg of this 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. (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 by Flow Cytometry Analysis: Mouse Strain: BALB/c Cell Concentration: 1x10e6 cells per test Antibody Concentration Used: 0.5 μg/10e6 cells Isotypic Control: PE Mouse IgG2a Percentage of cells stained above control: Thymus 86.3% Spleen 97.2% Lymph Node 100%
Reference Data

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