Rabbit anti pchk1 s345

Mouse anti-MBP (1:5000, R29, Cat. #MA5-14122, Thermo Fisher Scientific), mouse anti-BRCA2 (1:1000, OP95, EMD Millipore), mouse anti-CHK1 (1:1000, Cat. #2360, Cell Signaling Technology), rabbit anti-pCHK1-S345 (1:500, Cat #2348, Cell Signaling Technology), Horseradish peroxidase (HRP) conjugated secondary antibodies used: mouse-IgGκ BP-HRP (IB: 1:5000, Cat. #sc-516102, Santa Cruz), HRP Goat anti-mouse IgG (1:10,000, Cat #115-035-003, Jackson Immuno), HRP Goat anti-rabbit IgG (1:10,000, Cat #111-035-003, Jackson Immuno).

Chromatin fractionation experiments were performed as previously described (23 (link)). Western blotting was performed using standard methods. Blots were incubated with primary antibodies against: rabbit anti-pCHK1(S345) (Cell Signalling Technology), mouse anti-CHK1 (Santa Cruz Biotechnology), rabbit anti-RAD51 (Bioss Antibodies), mouse anti-RPA32 (Calbiochem), rabbit anti-RPA70 (GeneTex), mouse anti-GAPDH (Millipore) and rabbit anti-Lamin B1 (Abcam). After incubations with horseradish peroxidase-linked secondary antibodies (1:20 000, Jackson Immunosciences), the blots were developed using the chemiluminescence detection kit WesternBright ECL HRP substrate (Advansta) according to the manufacturer's instructions. Quantification was performed on scanned images of blots using the Image Lab software, and the values shown on the graphs represent normalization of the protein content evaluated through LaminB1 or GAPDH immunoblotting.