Antibody was affinity purified using an epitope specific to MED12 immobilized on solid support. The epitope recognized by A300-774A maps to a region between residue 2150 and the C-terminus (residue 2212) of human Mediator of RNA Polymerase II Transcription, subunit 12 using the numbering given in entry NP_005111.1 (GeneID 9968). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
activator-recruited cofactor 240 kDa component, ARC240, CAG repeat protein 45, CAGH45, FGS1, HOPA, human opposite paired, MED12S, Mediator complex subunit 12, mediator of RNA polymerase II transcription subunit 12, mediator of RNA polymerase II transcript
between 2127 and C-term
2 - 8°C
Detection of human and mouse Med12 by western blot. Samples: Whole cell lysate (50 µg) from HeLa and mouse NIH 3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-Med12 antibody A300-774A (lot A300-774A-2) used for WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Detection of human Med12 by western blot of immunoprecipitates. Samples: Whole cell lysate (0.5 or 1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-Med12 antibody A300-774A (lot A300-774A-2) used for IP at 6 µg per reaction. Med12 was also immunoprecipitated by a previous lot of this antibody (lot A300-774A-1). For blotting immunoprecipitated Med12, A300-774A was used at 0.4 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Detection of human MED12 by immunohistochemistry. Sample: FFPE section of human ovarian carcinoma. Antibody: Affinity purified rabbit anti-MED12 (Cat. No. A300-774A Lot2) used at a dilution of 1:1,000 (1µg/ml). Detection: DAB