United Kingdom (UK)
Recombinant monoclonal antibody to RT1-D. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma OX-17.
UniProt Accession Number of Target Protein: O98172
Alternative Name(s) of Target: MHC class II RT1D alpha chain antigen; OX17; MRC OX-17; MRC OX17
Immunogen: OX-17 was generated using BALB/c mice raised against rat spleen glycoproteins depleted of the Ia-A antigen using an OX-6 immunoaffinity column.
Specificity: The OX-17 antigenic determinant is isotypic and on the alpha chain. OX-17 labels B but not T cells from lymph node or thoracic duct lymph but very weakly labels thymocytes and does not label erythrocytes. OX-17 does not cross-react with Rat Ia-A or Mouse Ia-E antigen. The target antigen is expressed on antigen presenting cells, including B cells, macrophages and dendritic cells. RT1-D is a rat MHC class II protein. RT1-D is equivalent to mouse I-E antigen.
Application Notes: This antibody binds to rat MHC Class I, and has been used to show that, in an in vivo rat model, antibodies against MHC class I antigens from stored blood may precipitate transfusion-related acute lung injury (Kelher, 2009).
Antibody first published in:
Fukumoto et al Mouse monoclonal antibodies against rat major histocompatibility antigens. Two Ia antigens and expression of Ia and class I antigens in rat thymus. Eur J Immunol 12:237-43 19826456222 PMID:6178598
Note on publication:
Describes the generation and use of OX-17 to identify and purify novel Ia antigens..
Flow-cytometry using the anti-RT1-D antibody OX-17 (Ab00582) Rat splenocytes were fixed using 2% PFA, permeabilised using 0.5% Triton and stained with unimmunized rabbit IgG antibody (MOPC-21; isotype control, black line) or the rabbit IgG-chimeric version of OX-17 (Ab00582-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.
Immunofluorescence staining of fixed rat splenocytes with anti-RT1-D antibody OX-17 (Ab00582) Immunofluorescence analysis of paraformaldehyde fixed rat splenocytes permeabilized with 0.15% Triton and stained with the chimeric mouse IgG1 version of OX-17 (Ab00582-23.0) at 10 µg/ml for 1h followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab00582-23.0, DAPI, merged channels and an isotype control. The isotype control was stained with an anti-unknown specificity antibody (Ab178-23.0) followed by Alexa Fluor® 488 secondary antibody.
Western Blot using anti-RT1-D antibody OX-17 (Ab00582) Rat lymph node lysate (35µg protein in RIPA buffer) was resolved on a 10% SDS PAGE gel and blots probed with the chimeric rabbit IgG version of OX-17 (Ab00582-23.0) at 2 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence. The expected running size for RT1-D is 30.2kDa.