The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IP: Use at 5-10 µg/mg of lysate.
WB: 1/2000 - 1/10000. Predicted molecular weight: 233 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Caspase-like protease, which plays a central role in the chromosome segregation by cleaving the SCC1/RAD21 subunit of the cohesin complex at the onset of anaphase. During most of the cell cycle, it is inactivated by different mechanisms.
Belongs to the peptidase C50 family.
Autocleaves. This function, which is not essential for its protease activity, is unknown. Phosphorylated by CDK1. There are 8 Ser/Thr phosphorylation sites. Among them, Ser-1126 phosphorylation is the major site, which conducts to the enzyme inactivation.
Separase was immunoprecipitated using ab85685 at 10 µg/mg whole cell lysate from HeLa cells.
20% of IP was loaded per lane, and probed with ab85685 at 1 µg/ml (lane 1) or with a control IgG (lane 2).
Detection: chemiluminescence with exposure time of 10 seconds.
Note: The more rapidly migrating band (MW ~175kDa) is believed to represent an additional form of Separase
owing to recognition by multiple antibodies each of which was directed toward distinctly different epitopes on Separase.