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

SKU CL129BX

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

Product Data

• Clone Name: OX-27
• Application: FC, IHC
• Recommended Dilution: Flow Cytometry.; Immunohistochemistry using cryostat sections: (however, cross-reactivity with Lewis rats have been shown to occur in some instances).
• Reactivity: Rat
• Antibody Host: Mouse
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: Phytohaemagglutinin Blasts; Donor: BALB/c spleen; Fusion Partner: P3-NS1-1-Ag4 (NS1/1)
• Specificity: This Antibody recognizes a polymorphic determinant of the MHC class I antigen in the rat. (c and n haplotypes). This antibody is an excellent antibody for labelling cells of donor or host origin in bone marrow chimeras.
• Buffer: 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 fraction
• Concentration: lot specific
• Purification: Protein A Chromatography
• Conjugation: Biotin
• Storage: Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.; Avoid repeated freezing and thawing.
• Stability: Shelf life: one year from despatch.
• Shipping: Blue Ice
• Database Link:
  Q31255
• 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: 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 0.5-1.0 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. Not recommended for use with a PE secondary.; 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: Brown Norway; Cell Concentration: 1x10e6 cells per test; Antibody Concentration Used: 1.0 µg/10e6 cells; Isotypic Control: Biotin Mouse IgG2a; ; Cell Source Percentage of cells stained above control:; Thymus 65.4%; Spleen 98.9%; Lymph Node 100%; ; Restults - Strain Distribution:; Cell Concentration: 1x10e6 cells per test; Antibody Concentration Used: 1.0 µg/10e6 cells; Strains Tested: Brown Norway, Buffalo, Wistar, Fischer, ACI; Positive: Brown Norway; Negative: Buffalo, Wistar, Fischer, ACI

Reference Data