Clinisciences > MHC Class I (RT1Aa) mouse monoclonal antibody, clone OX-18, Azide Free
MHC Class I (RT1Aa) mouse monoclonal antibody, clone OX-18, Azide Free
Referencia SM311A
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MHC Class I (RT1Aa) Mouse Monoclonal Antibody [Clone ID: OX-18]
| 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 | |
| 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 |
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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.