PARP-1 monoclonal antibody (F1-23)

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	-Mouse (1:1000)

Protocol tips
-Mouse (1:1000)

Publication protocol

Cells were lysed in sample buffer (125 mM Tris–HCl pH 6.8, 20% Glycerol, 10% 2-β-mercaptoethanol, 4% SDS, 0.01% Bromophenol Blue) and boiled for 5 min at 98°C. Proteins were separated in SDS-PAGE gels and transferred onto PVDF membranes (0.45 μm, Merck Millipore). After 1 h of blocking in 5% BSA in PBS-T (0.05% Tween 20), membranes were incubated with primary antibodies in PBS-T at 4°C overnight. Antibodies were against CSA (EPR9237, 1:1000, Abcam, ab137033), CSB (E-18, 1:250, Santa Cruz, sc-10459), XPF (3F2/3, 1:500, Santa Cruz, sc-136153), XPA (FL-273, 1:250, Santa Cruz, sc-853), POLB (1:2000, Abcam, ab26343), PARP1 (F1-23, 1:1000, Alexis Biochemicals, ALX-804-211), XRCC1 (C-15, 1:500, Santa Cruz, sc-5903), OGG1 (EPR4664(2), 1:1000, Abcam, ab124741) and, as loading control, against Ku70 (M-19, 1:1000, Santa Cruz, sc-1487), Tubulin (B-512, 1:5000, Sigma-Aldrich, T6074) and Aquarius (1:1000, Bethyl Laboratories, A302-547A). Anti-mouse, anti-goat and anti-rabbit secondary antibodies, conjugated to CF™680 or CF™770 dyes (Sigma-Aldrich) were used at a dilution of 1:10 000. Probed membranes were visualized with the Odyssey CLx Infrared Imaging System (LI-COR Biosciences).

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Manufacturer protocol

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