United Kingdom (UK)
Recombinant monoclonal antibody to TCR. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma H57-597.
UniProt Accession Number of Target Protein: P01852
Alternative Name(s) of Target: TCR-beta; T cell receptor; T-cell receptor; T cell receptor beta chain; TCRB; TRB; TCB1; Ti
Immunogen: Hamsters were immunised with affinity purified DO-11.10 TCR, and produced detectable anti-TCR antibodies which were judged by the ability of the anti-sera to immunoprecipitate TCR from surface labelled DO-11.10 T cell hybridomas. Hamster 57 was selected for hybridisation, and B cell hybridomas were produced using polyethylene glycol fusion of hamster splenic B cells to the mouse myeloma variant. Selected mAb were screened after subsequent culturing for their ability to immunoprecipitate TCR from detergent lysates of surface-radiolabelled murine T cell hybridomas.
Specificity: mAb H57-597 reacts with surface receptors on all alpha-beta TCR bearing cells, but does not react with receptors on gamma-delta T cells. H57-597 is directed to an epitope within the FG loop of the C region of TCR beta chain.
Application Notes: In an immobilised form, this antibody can directly activate T cells bearing alpha-beta TCR to produce IL-2. H57-597 has also been used to construct a Fab fragment which has been complexed with deglycosylated TCRs for their crystallisation for structure determination (Liu 1996). Flow cytometry is another application of this antibody, where H57-597 has been used to analyse TCR-beta expression in different T cell populations (Tsukumo 2006).
Antibody first published in:
Kubo et al. Characterisation of a monoclonal antibody which detects all murine alpha-beta T cell receptors J Immunol. 1989 Apr 15;142(8):2736-42. PMID:2467936
Note on publication:
Describes the need for a monoclonal antibody that detects alpha-beta but not delta-gamma TCRs, to allow enumeration of the two types of cells separately during development. The generation of the antibody is then outlined.
Immunofluorescence staining of mouse splenocytes with anti-TCR (Ab00903) H57-597 Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of H57-597 (Ab00903-23.0) 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 Ab00903-23.0, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.
Flow cytometry using the Anti-TCR antibody H57-597 (Ab00903). Paraformaldehyde fixed mouse splenocytes were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of H57-597 (Ab00903-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the 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 mouse splenocytes with anti-TCR (Ab00903) H57-597 Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of H57-597 (Ab00903-23.0) 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 Ab00903-23.0, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.