Antibody was affinity purified using an epitope specific to BID phosphorylated on Serine 78 immobilized on solid support. The epitope recognized by A300-528A maps to phosphorylated Serine 78 and surrounding residues of mouse BH3 interacting domain death agonist using the numbering given in entry NP_031570.2 (GeneID 12122). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
2700049M22Rik, AI875481, AU022477, BH3-interacting domain death agonist, BID, p22 BID
surrounding serine 78
2 - 8°C
Detection of mouse Phospho-BID (S78) after Treatment with Etoposide. Samples: Whole cell lysate (50 and 80 µg for WB; 1 mg for IP, 20% of IP loaded) from NIH 3T3 cells treated with 100 µM etoposide for 3 hours (+) or mock treated (-) cells. Antibodies: Affinity purified rabbit anti-phospho BID (S78) antibody A300-528A (lot A300-528A-2) used for WB at 1 µg/ml (upper panels of A and B). Rabbit anti-BID antibody BL1079 was used for IP of BID and phospho-BID and for detection of total BID (lower panels of A and B). Detection: Chemiluminescence with exposure times of 3 minutes (upper panels of A and B).