This affinity purified antibody is directed against the phosphorylated form of human APC1 at the pS355 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide.
This is one of a number of novel phosphorylation sites for APC1 identified in Kraft C et al. It may be involved in the regulation of the APC/C.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/200 - 1/2000. Detects a band of approximately 215 kDa (predicted molecular weight: 215 kDa).
1/10000 - 1/40000.
Tested in peptide ELISA. Product has been tested against 0.1 µg of the phosphorylated and non-phosphorylated forms of the immunizing peptide.
Component of the anaphase promoting complex/cyclosome (APC/C), a cell cycle-regulated E3 ubiquitin ligase that controls progression through mitosis and the G1 phase of the cell cycle. The APC/C complex acts by mediating ubiquitination and subsequent degradation of target proteins: it mainly mediates the formation of 'Lys-11'-linked polyubiquitin chains and, to a lower extent, the formation of 'Lys-48'- and 'Lys-63'-linked polyubiquitin chains.
Protein modification; protein ubiquitination.
Belongs to the APC1 family. Contains 4 PC repeats.
Phosphorylated. Phosphorylation on Ser-355 occurs specifically during mitosis.
Western blot - Anti-Apc1 (phospho S355) antibody (ab10923)
All lanes : Anti-Apc1 (phospho S355) antibody (ab10923) at 1/500 dilution
Lane 1 : HeLa S3 whole cell lysate (asynchronous cells) Lane 2 : HeLa S3 whole cell lysate (cells synchronized at G1/S boundary by treatment with thymidine) Lane 3 : HeLa S3 whole cell lysate (cells synchronized at M phase by treatment with nocodazole)
Lysates/proteins at 30 µg per lane.
Secondary All lanes : Goat anti-Rabbit IgG [H&L] at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 215 kDa Observed band size: 215 kDa
Phosphorylated APC1 is mostly present only in cell preparations arrested at cell division.