MHC Class I (RT1Ac) Mouse Monoclonal Antibody [Clone ID: OX-27]

CAT#: CL129FX

MHC Class I (RT1Ac) mouse monoclonal antibody, clone OX-27, FITC



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CNY 10,350.00


货期*
5周

规格
    • 500 ug

Product images

Specifications

Product Data
Clone Name OX-27
Applications FC
Recommend Dilution Flow cytometry: use 0.5 µg of neat antibody to label 10e6 cells.
Reactivity Rat
Host Mouse
Clonality Monoclonal
Immunogen Phytohaemagglutinin Blasts.
Formulation PBS buffer with 0.02% sodium azide as preservative and 0.5 % EIA grade BSA as stabilizer.
Label: FITC
State: Liquid purified IgG fraction.
Label: conjugated
Concentration lot specific
Purification Protein G Chromatography.
Conjugation FITC
Storage Condition Store the antibody at 2-8°C for one month or (in aliquots) at -20°C for longer.
Avoid repeated freezing and thawing.
Background MHC Class I molecules play a central role in the immune system by presenting peptides derived from the endoplasmic reticulum lumen. MHC class I antigens are heterodimers consisting of one alpha chain (44kDa) with beta 2 microglobulin (11.5 kDa). The antigen is expressed by all somatic cells at varying levels. MHC Class I molecules are expressed on most nucleated cells where they present endogenously synthesized antigenic peptides to CD8+ T lymphocytes, which are usually cytotoxic T cells. Fibroblasts or neurons however only show a low level of antigen.
Synonyms RT1-A3
Note Recognizes a polymorphic determinant of the MHC class I antigen in the rat.
This Antibody can be used for labelling cells of donor or host origin in bone marrow chimeras. (1,2)

Protocol: FLOW CYTOMETRY ANALYSIS:

Method:
1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with 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 0.5-0.2 µg antibody 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. (protect tubes from light)
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:
Rat Strain: Brown Norway
Cell Concentration: 1x10e6 cells per test.
Antibody Concentration Used: 0.2 µg/10e6 cells
Isotypic Control: FITC Mouse IgG2a.

Cell Source-Percentage of cells stained above control:
Thymus: 73.9%
Spleen: 99.0%
Lymph Node: 100%
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.
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