United Kingdom (UK)
Recombinant monoclonal antibody to Rad51D. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 5A8/4.
UniProt Accession Number of Target Protein: O75771
Alternative Name(s) of Target: DNA repair protein RAD51 homolog 4; R51H3; RAD51 homolog D; RAD51-like protein 3; TRAD; HsTRAD; R51H3Trad; RAD51DRAD51 (S. cerevisiae)-like 3; RAD51-like 3 (S. cerevisiae); recombination repair protein; RAD51L3; 5A8
Immunogen: This antibody was raised by immunising mice with the denaturated recombinant human Rad51D protein which had been previously amplified and purified from E. coli.
Specificity: This antibody recognises human and hamster Rad51D. Rad51D, paralog of Rad51 which is an E. coli RecA homologue. It is invloved in the homologous recombination repair (HRR) pathway which fixes double-stranded breaks.RAD51 forms a nucleoprotein filament which initiates recombination.
Application Notes: WB: Rad51D 5A8/4 (5A8) antibody detected Rad51D in a material from V79 and irs3 cells in Western blot analysis of immunoprecipitated Rad51 complexes (French et al., 2002). WB: This antibody was also applied in experiments studying the role of Rad51D in the DNA repair where the antibody detected Rad51D protein on Western blots from CHO AA8 cells (Hinz et al., 2006).
Antibody first published in: French et al. Role of mammalian RAD51L2 (RAD51C) in recombination and genetic stability. PMID:11912211 Note on publication: This article describes the earliest use of Rad51D 5A8/4 (5A8) antibody to detect Rad51D protein via Western blotting. However, the exact process of generation of this antibody is described in the following paper (where similar antibodies were generated): Masson, J. Y., Stasiak, A. Z., Stasiak, A., Benson, F. E., & West, S. C. (2001). Complex formation by the human RAD51C and XRCC3 recombination repair proteins. Proceedings of the National Academy of Sciences, 98(15), 8440-8446.
Immunofluorescence staining of HeLa cells using anti-Rad51D (Ab01307) 5A8/4 Immunofluorescence analysis of paraformaldehyde fixed HeLa cells stained with the chimeric mouse IgG version of 5A8/4 (Ab01307-1.1) at 10 µg/ml followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing nuclear staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab01215-1.1, DAPI, merged channels and an isotype control. The isotype control was stained with anti-unknown antibody (Ab00178-1.1) followed by Alexa Fluor® 488 secondary antibody.
Flow cytometry using the Anti-Rad51D antibody 5A8/4 (Ab01307). Paraformaldehyde fixed HeLa cells permeabilized with 0.5% Triton were stained with anti-unknown specificity antibody (Ab00178-1.1; isotype control, black line) or the mouse IgG1 version of 5A8/4 (Ab01307-1.1, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-mouse IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.