MHC Class II I-Ap Mouse Monoclonal Antibody [Clone ID: 7-16.17]

Specifications

Product Data
Clone Name	7-16.17
Applications	FC
Recommend Dilution	Flow Cytometry.
Reactivity	Mouse
Host	Mouse
Clonality	Monoclonal
Immunogen	B10.p Donor: BALB/c Fusion Partner: SP2/0
Specificity	This monoclonal antibody is a cytotoxic antibody which defines a public I-A antigen. This antibody reacts with I-A antigen from the following I-A haplotypes: I-Ap,k,q,r,s,b. Using recombinant strains, reactivity against the b haplotype has been localized to the Ab subregion. This antibody can be used to quantitate or eliminate I-A bearing cells for precipitating I-A antigen.
Formulation	PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml. Label: FITC State: Liquid purified
Concentration	lot specific
Purification	Protein G Chromatography
Conjugation	FITC
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. This product is photosensitive and should protected from light.
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 0.1 - 0.2 µg* of this Ab 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: BDP Cell Concentration: 1x10e6 cells per tests Antibody Concentration Used: 0.1 µg/10e6 cells Isotypic Control: FITC Mouse IgG2a Cell Source - Percentage of cells stained above control: Spleen: 76.7% Lymph Node: 40.5% Bone Marrow: 39.4% Thymus: 55.6% Strain Distribution by Flow Cytometry Analysis: Procedure: As above Antibody Concentration: 0.2 µg/10e6 cells Strains Tested: see FIGURE 2 For a more detailed strain distribution - see reference 1.
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.