MHC Class II (I-Ab,d) Mouse Monoclonal Antibody [Clone ID: 25-9-17S]

Specifications

Product Data
Clone Name	25-9-17S
Applications	FC
Recommend Dilution	Flow Cytometry (See Protocols).
Reactivity	Human
Host	Mouse
Clonality	Monoclonal
Immunogen	C3H.SW splenocytes. Donor: C3H lymphoid cells. Fusion Partner: Sp2/0-Ag14
Specificity	This Monoclonal Antibody reacts with I-Ab and I-Ad antigens. Cross reaction with H-2p and H-2q was also found.
Formulation	PBS containing 0.02% Sodium Azideas preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml Label: Biotin State: Liquid purified Ig fraction
Concentration	lot specific
Purification	Protein G Chromatography
Conjugation	Biotin
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.
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 106 cells, representing 1 test). 4. To each tube, add 0.5-0.2 μg* of AM31877BT-N 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. Add 100 μl of secondary antibody (Streptavidin-FITC) at a 1:500 dilution. 9. Incubate tubes at 4°C for 30-60 minutes (It is recommended that tubes are protected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C. 11. Resuspend the cell pellet in 50 μl ice cold media B. 12. 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: C57BL/6 Cell Concentration : 1x10e6 cells per test. Antibody Concentration Used: 0.5 μg/106 cells. Isotypic Control: Biotin Mouse IgG2a Cell Source Percentage of cells stained above control: Thymus: 17.5% Spleen: 58.6% Lymph Node: 31.2% Strain Distribution by Flow Cytometry Analysis: Antibody Concentration: 0.5 µg/106 cells. Strains Tested: See Figure 2.
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.