MHC Class I (RT1Aa) Mouse Monoclonal Antibody [Clone ID: OX-18]

SKU SM311A

MHC Class I (RT1Aa) mouse monoclonal antibody, clone OX-18, Azide Free

Product Data

• Clone Name: OX-18
• Application: FC, IHC, IP
• Recommended Dilution: Flow Cytometry (see Protocols)
• Reactivity: Rat
• Antibody Host: Mouse
• Isotype: IgG1
• Clonality: Monoclonal
• Immunogen: Rat spleen membrane glycoproteins depleted of Ia-A antigens.; Donor: BALB/c spleen.; Fusion Partner: X63 Ag8.653.
• Specificity: This monoclonal antibody recognizes a monomorphic determinant of rat Class I MHC antigen (RT1.A) and thus reacts with all rat strains tested including AO(RT1µ), DA (RT1a), LEW (RT11) and PVG (RT1c). However, quantitiative measurements suggest that only a subfraction of the total Class I molecules are recognized (2). This antibody labels all peripheral lymphocytes but only a subfraction of thymocytes. It may be that the weakly labelled cells do in fact express some OX-18 antigenic determinants but there is clearly a major quantitative difference amongst thymocytes. This monoclonal used in tissue sections preferentially labels lymphoid cells in the medulla of the thymus including those cells with the marker phenotype of mature T lymphocytes.
• Buffer: PBS; State: Azide Free; State: Liquid pruified Ig
• Concentration: lot specific
• Purification: Protein G Chromotraphy
• Conjugation: Unconjugated
• 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.; Handle under aseptic conditions.
• Stability: Shelf life: one year from despatch.
• Shipping: Blue Ice
• Database Link:
  P16391
• 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.
• 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 50µl of this suspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test).; 4. To each tube, add 0.2-0.5 µg* of this Ab.; 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 (Goat anti-mouse IgG (H+L)-FITC conjugate) at 1:500 dilution.; 9. Incubate tubes at 4°C for 30-60 minutes. (It is recommended that the 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: Wistar; Cell Concentration: 1x10e6 cells per test; Antibody Concentration Used: 0.2 µg/10e6 cells; Isotypic Control: Mouse IgG1,k; ; Cell Source Percentage of cells stained above control:; Thymus: 38.1%; Spleen: 97.0%; Lymph Node: 99.9%; ; Results - Strain Distribution:; Antibody Concentration Used: 0.2 µg/10e6 cells; Strains Tested: Wistar, Buffalo, Fischer 344; Positive: Wistar, Buffalo, Fischer 344; Negative: None

Reference Data