United Kingdom (UK)
Recombinant monoclonal antibody to MHC I. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma R1-21.2.
Alternative Name(s) of Target: R1.21.2; H-2; MHC-I; MHC Class I
Immunogen: This antibody was raised by immunising Lewis 1F rats with mouse CH-1 lymphoma cells. Spleen cells from immunised rats were fused with X-63Ag8-653 myeloma cells to generate hybridomas.
Specificity: This antibody is specific for murine MHC class I, and binds to polymorphic H-2 determinants expressed on most haplotypes. This antibody is, therefore, able to bind all H-2 class I molecules, independent of the allele. This antibody inhibits the binding of most mouse anti-H-2 antibodies to H-2k cells.
Application Notes: This antibody binds murine H-2 MHC class I molecules, and has been used in both structural and biochemical studies. It has been employed to assess MHC class I expression in frozen, acetone-fixed mouse brain sections by immunohistochemistry (Miralvès et al, 2007), and to stain paraffin-embedded sections of mouse liver for H-2Kb expression (Limmer et al, 2005). It has also been used to precipitate mouse H-2Kk and H-2Dk class I molecules from detergent lysates of Ltk3.3/1 cells (Rada et al, 1990), and to stain H-2Kb and H-2Kk-expressing cells in FACS analysis (Bendelac, 1995). Cellular radioimmune assays have been used to confirm the binding of this antibody to spleen cells, and assess its strain specificity (Koch et al, 1983).
Antibody first published in:
Koch et al Comparison of allogeneic and xenogeneic determinants on the H-2Kk molecule Transplantation, August 1983, Vol.36(2), pp.177-80 [Peer Reviewed Journal] PMID:6192569
Note on publication:
Describes the original generation and characterisation of this antibody.
Immunofluorescence staining of mouse splenocytes using anti-MHC I antibody (Ab00880) R1-21.2. Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes immobilized on Shi-fix™ cover-slips and stained with the chimeric rabbit IgG version of R1-21.2 (Ab00880-23.0) at 10 µg/ml followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining in a subset of cells. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab00880-23.0, DAPI, merged channels and an isotype control. The isotype control was stained with anti-Fluorescein antibody (Ab00102-23.0) followed by Alexa Fluor® 488 secondary antibody.
Western Blot using anti-MHC I antibody R1-21.2 (Ab00880) Mouse thymus tissue lysate (35µg protein in RIPA buffer) was resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of R1-21.2 (Ab00880-23.0) at 1 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.