MHC Class II I-Ak Mouse Monoclonal Antibody [Clone ID: 14V.18]

CAT#: SM083B

MHC Class II I-Ak mouse monoclonal antibody, clone 14V.18, Biotin


USD 250.00

2 Weeks*

Size
    • 100 ug

Product Images

Specifications

Product Data
Clone Name 14V.18
Applications Assay, FC
Recommended Dilution Flow Cytomtry.
Reactivities Mouse
Host Mouse
Isotype IgG2a
Clonality Monoclonal
Immunogen A.TL spleen
Donor: A.TH
Fusion Partner: P3-X63-Ag 8
Specificity This monoclonal antibody is a cytotoxic antibody specific for cells expressing the Ia antigen coded for by the A subregion of the k haplotype. The reaction pattern of this antibody with a panel of inbred and recombinant haplotypes demonstrates that the antibody reacts with Ia.m2, a private specificity of the H-2k haplotype. This antibody can be used to quantitate or to eliminate cells bearing the I-Ak (Ia.m2) 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: Biotin
State: Liquid purified Ig
Concentration 0.1 mg/ml
Purification Protein G Chromatography
Conjugation Biotin
Synonyms H2-Aa
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 1.0 - 0.5 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.
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:
Mouse Strain: A.TL
Cell Concentration: 1x10e6 cells per tests
Antibody Concentration Used: 0.5 µg/10e6 cells
Isotypic Control: Biotin Mouse IgG2a

Cell Source Percentage of cells stained above control:
Thymus: 23.6%
Spleen: 63.9%
Lymph Node: 32.1%
Bone Marrow: 11.1%

Results - Strain Distribution:
Antibody Concentration Used: 1.0 µg/10e6 cells
Strains Tested: see Picture 1
Reference Data

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*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.