United Kingdom (UK)
Recombinant monoclonal antibody to Rad54. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 4E3/1.
UniProt Accession Number of Target Protein: Q92698
Alternative Name(s) of Target: DNA repair and recombination protein RAD54-like; RAD54 homolog; hHR54; hRAD54; RAD54ADNA repair and recombination protein RAD54-like; RAD54-like (S. cerevisiae); hRAD54HRAD54; HR54; EC 3.6.1; hHR54EC 3.6.4.-; RAD54 like
Immunogen: This antibody was raised against full length human Rad54 protein.
Specificity: This antibody recognises human Rad54. Rad54 is a protein engaged in recombination during mitosis as well as DNA repair mechanisms.
Application Notes: IF and IP: This antibody was used in an immunoprecipitation experiment and to investigate colocalisation of Rad54 with S100A11 via immunofluorescence in fixed HaCaT cells treated with BLM (Murzik et al., 2008). WB and IF: Rad51 was visualised both on Western blots and immunofluorescently in BS/A-15 and NHF cells with this anti-Rad54 antibody (Srivastava et al., 2009).
Antibody first published in:
Murzik et al. Rad54B targeting to DNA double-strand break repair sites requires complex formation with S100A11. Mol Biol Cell. 2008 Jul;19(7):2926-35. doi: 10.1091/mbc.E07-11-1167. Epub 2008 May 7. PMID:18463164
Note on publication:
This article describes the earliest identified use of the anti-Rad54 Rad54 4E3/1 antibody. It was used to investigate the potential interaction and colocalisation between Rad54 and another protein (IP and IF experiments, supplemental materials of the article).
Immunofluorescence staining of HeLa cells using anti-Rad54 antibody (Ab01309) 4E3/1. Immunofluorescence analysis of paraformaldehyde fixed HeLa cells permeabilized with 0.15% Triton and stained with the chimeric mouse IgG version of 4E3/1 (Ab01309-1.1) at 10 µg/ml followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing nulcear staining. Actin filaments were stained with phalloidin (red) and the nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab01309-1.1, DAPI, merged channels and an isotype control. The isotype control was stained with anti-unknown antibody (Ab00178-1.1) followed by Alexa Fluor® 488
Flow-cytometry using the anti-Rad54 antibody 4E3/1 (Ab01309). HeLa cells were fixed using paraformaldehyde, permeabilised using 0.5% Triton and stained with anti-unknown IgG antibody (Ab00178-1.1; isotype control, black line) or the mouse IgG-chimeric version of 4E3/1 (Ab01309-1.1, blue line) at a dilution of 1:100 for 1h at RT. After washing, 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.
Western Blot using anti-Rad54 antibody 4E3/1 (Ab01309). nuclear lysates of HEK293(A), HeLa(B), Jurkat(C) and K562(D) cells (35µg protein in RIPA buffer) was resolved on an SDS PAGE gel and blots probed with the chimeric mouse IgG version of 4E3/1 (Ab01309-1.1) at 2 µg/ml before detection using an anti-mouse secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Flow cytometry using the Anti-Rad54 antibody 4E3/1 (Ab01309). 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 4E3/1 (Ab01309-1.1, 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.