MHC Class II RT1D Mouse Monoclonal Antibody [Clone ID: OX-17]

CAT#: CL121BX

MHC Class II RT1D mouse monoclonal antibody, clone OX-17, Biotin


USD 505.00

2 Weeks*

Size
    • 500 ug

Product Images

Specifications

Product Data
Clone Name OX-17
Applications FC, IHC
Recommended Dilution Flow Cytometry.
Reactivities Rat
Host Mouse
Isotype IgG1
Clonality Monoclonal
Immunogen Rat spleen membrane glycoproteins depleted of Ia-A antigens.
Immunocyte Donor: BALB/c spleen
Fusion Partner: X63 Ag8.653
Specificity This monoclonal antibody recognizes a monomorphic determinant on the a chain of the rat Ia antigen and appears to be the rat homologue of mouse Ia-E. It recognizes the rat Ia product present on B, but not T cells from lymph node or thoracic duct lymph. It does not bind to thymocytes or erythrocytes. The antibody does not cross-react with rat Ia-A or mouse Ia-E antigen, but rabbit antibody raised against the antibody affinity column-purified MRC OX-17 antigen cross-reacted on tissues of mice expressing Ia-E mouse antigen but not on those mouse strains that were Ia-E antigen negative.
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 HLA Class II
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®-Rat 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 0.5-0.1 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:
Rat Strain: Fischer
Cell Concentration: 1x10e6 cells per test
Antibody Concentration Used: 0.1 µg/10e6 cells
Isotypic Control: Biotin Mouse IgG1

Cell Source Percentage of cells stained above control:
Thymus 6.1%
Spleen 48.8%
Lymph Node 27.5%
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.