MHC Class I H2 Kd/Dd Mouse Monoclonal Antibody [Clone ID: 34-1-2S]

CAT#: AM08081BT-S

MHC Class I H2 Kd/Dd mouse monoclonal antibody, clone 34-1-2S, Biotin


USD 250.00

2 Weeks*

Size
    • 100 ug

Product Images

Specifications

Product Data
Clone Name 34-1-2S
Applications Assay, FC
Recommended Dilution Flow Cytometry (See Protocols for more details).
Results:
Tissue Distribution by Flow Cytometry Analysis:
Mouse Strain: BALB/c
Cell Concentration : 1x10e6 cells per test
Antibody Concentration Used: 0.5 µg/10e6 cells.
Isotypic Control: Biotin Mouse IgG2a
Cell Source: Percentage of cells stained above control
Thymus: 82.5%
Spleen: 95.7%
Lymph Node: 100%
Reactivities Mouse
Host Mouse
Isotype IgG2a
Clonality Monoclonal
Immunogen BDF splenocytes.
Donor: C3H spleen.
Fusion Partner: Sp2/0-Ag14.
Specificity This Monoclonal Antibody reacts with both H-2Kd and H-2Dd products. The antibody also cross reacts with Kbsrpq.
This K-D cross reaction indicates the presence of shared specificities between the two separate H-2 regions.
Formulation PBS containing 0.02% Sodium Azide as 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 0.1 mg/ml
Purification Protein G Chromatography
Conjugation Biotin
Background The 'classical' MHC Class I molecules are histocompatibility antigens encoded by the H-2 gene complex and consist of heterodimers of highly polymorphic alpha chains noncovalently associated with the invariant beta 2-Microglobulin. (Ref.3,4) These antigens are expressed on most nucleated cells but expression varies on different cell types. MHC Class I molecules present endogenously synthesized peptides to CD8+ T lymphocytes, which are usually cytotoxic T cells. (Ref.5) MHC Class I antigens expressed on thymic epithelial cells regulate the positive and negative selection of CD8+ T cells during T cell ontogeny. (Ref.3,6)
Note Strain Distribution by Flow Cytometry Analysis:
Antibody Concentration: 0.2 µg/106 cells.
Strains Tested (Figure 2): See Ref.9 for a more detailed strain distribution.

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 2x107 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 AM08081BT-S 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).
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.