Anti-Rad51 抗体 (ab176458)

Immunofluorescence detection of Rad51 foci formation after X-ray irradiation in GM0637 cells with ab176458 at 1/10000 dilution (left panels) and 1/1000 dilution (right panels).  The secondary antibody, anti-rabbit Alexa 488 was used at 1/10000 dilution.

Cells were irradiated by X-rays at 2 Gy, grown for 1 hr, fixed with 4% paraformaldehyde in 1x PBS for 10 min, washed 3 times with PBS for 3 min, permealized by treatment with 0.5% Triton for 5 min, washed 3 times with PBS for 3 min, incubated with ab176458 for 30 min at 37°C, washed 3 times with PBS for 3 min, incubated with secondary antibody for 30 min at 37°C, washed 3 times with PBS for 3 min, stained with Hoechst for 1 min and mounted.

The pictures were by courtesy of Prof. S. Tashiro and Dr. K. Kono at Hiroshima University.

ab176458 (20 µg) was incubated with 20 μg of HeLa cell extract, and precipitated with 20 μg of proteinA-beads. The sample was dissociated from the precipitate by heating in SDS-sample buffer and analyzed by western blotting with anti-Rad51 antiserum (chicken, ab63802) at 1/1000 dilution. As secondary antibody, anti-chicken IgG antibody (rabbit) was used at 1/10000 dilution.

Immunohistological staining of Rad51 protein in mouse testis using ab176458. A section of formalin fixed and paraffin embedded mouse testis was treated with ab176458 at 1/100 dilution after deparaffization and antigen retrieval. The secondary antibody, Alexa Fluor^® 647conjugated anti-rabbit IgG was used at 1/1,000 dilution (top left). The sample was counter-stained with DAPI (top right) and the merged image is shown (bottom left). The white light image of the same region is shown on the bottom right.