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

Specifications

Product Data
Clone Name	7-16.17
Applications	CT, FC
Recommend Dilution	Flow cytometry (protocol see below). Cytotoxic assay.
Reactivity	Mouse
Host	Mouse
Clonality	Monoclonal
Immunogen	B10.p
Specificity	CL072P is specific for Mouse-I-Ap,k,q,r,s,b. This antibody is a cytotoxic antibody which defines a public I-A antigen. Using recombinant strains, reactivity against the b haplotype has been localized to the Ab subregion. The antibody can be used to quantitate or eliminate I-A bearing cells for precipitating I-A antigen.
Formulation	PBS and 0.02% sodium azide (NaN3). State: Purified State: Liquid Ig fraction
Concentration	lot specific
Purification	Protein G chromatography
Conjugation	Unconjugated
Storage Condition	Store the antibody at 2 - 8 °C for up to one month. For long term storage, aliquot and freeze unused portion at -20 °C in volumes appropriate for single usage. 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. 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 0.2-1.0 µg of antibody. 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 (FITC Goat anti-mouse IgG) at 1/500 dilution. 9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes are protected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C in media B. 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). Strain Distribution by Flow Cytometry Analysis: Procedure: As above Antibody Concentration: 0.5 µg/10e6 cells Strains Tested: see picture below. For a more detailed strain distribution - see reference 1.
Reference Data

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