Ecl plus chemiluminescence system

Immunoprecipitation was performed as reported previously [13 (link)] by incubating the samples with rabbit polyclonal anti-MRE11 and anti-NBS1 antibodies (Novus) or anti-FXR1 rabbit polyclonal antibody (Bethyl). Co-immunoprecipitated proteins were detected by western blot analysis.
Western blotting of whole cell extract was performed as described previously [13 (link)]. Target proteins were detected with the primary antibodies listed above and secondary horseradish peroxidase-conjugated anti-rabbit or anti-mouse IgG (GE Healthcare) and visualized using the ECL plus chemiluminescence system (GE Healthcare).
Immunofluorescence staining was carried out as described previously [15 (link)] using Alexa-488-conjugated anti-rabbit or Alexa-594-conjugated anti-mouse IgG (Molecular Probes) to visualize the localization of target proteins.

Immunoprecipitation was performed as reported previously [13 (link)] by incubating the samples with monoclonal antibodies against γ-H2AX, RPA32, PCNA or RAD17. Co-immunoprecipitated proteins were detected by western blotting analysis.
Western blotting analysis of whole cell extract or chromatin fraction isolation was performed as described previously [10 (link), 13 (link)]. Target proteins were detected with the primary antibodies listed above and secondary horseradish peroxidase-conjugated anti-rabbit or anti-mouse IgG (GE Healthcare) and visualized using the ECL™ plus chemiluminescence system (GE Healthcare). Quantification of visualized bands was carried out by ImageJ software, and the ratios to unirradiated samples of negative knockdown were calculated [13 (link)].
Immunofluorescence staining was carried out as described previously [10 (link)] using Alexa-488–conjugated anti-rabbit or alexa-594–conjugated anti-mouse IgG (Molecular Probes) to visualize the localization of the target proteins.

Western blot analyses were carried out as described previously [12 (link)]. Target proteins were detected with the primary antibodies listed above and horseradish peroxidase (HRP)-conjugated anti-rabbit IgG or anti-mouse IgG antibodies (GE Healthcare). The bands of target proteins were visualized using the ECL plus chemiluminescence system (GE Healthcare). Quantification of the visualized bands was carried out by ImageJ software, and the ratios to quantities in unirradiated samples were calculated.