Antibody was affinity purified using an epitope specific to MDC1 immobilized on solid support. The epitope recognized by A300-051A maps to a region between residues 475 and 525 of human Mediator of DNA Damage Checkpoint 1 using the numbering given in entry NP_055456.1 (GeneID 9656). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
homologue to Drosophila photoreceptor protein calphotin, mediator of DNA damage checkpoint protein 1, NFBD1, nuclear factor with BRCT domains 1
between 475 and 525
2 - 8°C
Detection of human MDC1 by western blot. Samples: Whole cell lysate (15 and 50 µg) from HeLa cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-MDC1 antibody A300-051A (lot A300-051A-3) used for WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.
Detection of human MDC1 by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells. Antibodies: Affinity purified rabbit anti-MDC1 antibody A300-051A (lot A300-051A-3) used for IP at 3 µg per reaction. MDC1 was also immunoprecipitated by rabbit anti-MDC1 antibodies A300-052A and A300-053A. For blotting immunoprecipitated MDC1, A300-051A was used at 0.4 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.