2BN hTert (XLF-deficient) human fibroblasts were fixed for 30 min with methanol at −20°C, dipped for 1 min in ice cold acetone for permeabilization and washed three times for 10 min with PBS/1% FCS. Non-specific antigens were blocked for 30 min in 5% BSA in PBS/1% FCS. Samples were incubated with ab63801 (1∶15000) in PBS/1% FCS over night at 4°C, washed three times in PBS/1% FCS and incubated for 1 h at room temperature with AlexaFluor 488 conjugated secondary antibodies (1 : 500). After three times of washing in PBS, cells were DAPI stained.

Image shows 2BN hTert (XLF-deficient) human fibroblasts analyzed 2 h post IR with 1 Gy.

Anti-Rad51 antibody (ab63801) at 1/1000 dilution + Crude extract of Xenopus

Predicted band size: 37 kDa

Immunocytochemistry/ Immunofluorescence analysis of human osteosarcoma U2OS cells X-ray irradiation before and 2h after, immunostained with Anti-Rad51 antibody (ab63801) at 1/6,000 dilution. The lower panels were the same cells with Hoechst.

All lanes : Anti-Rad51 antibody (ab63801) at 1/2000 dilution

Lane 1 : Molecular weight markers
Lane 2 : HeLa cell extract

Predicted band size: 37 kDa
Observed band size: 37 kDa

Immunofluorescent detection of Rad51 foci formation induced by DNA damage. Normal human diploid cells were irradiated by X-ray (0.5 Gly) and after incubation of 6 hr, the cells were fixed and immuno-stained by using ab63801 (x100 dilution) as the primary antibody and Alexa594 labeled anti-rabbit antibody as the secondary antibody.