Western blot using ANAPC1 (phospho S377) polyclonal antibody (Cat # PAB9936) shows detection of a band ~215 KDa corresponding to phosphorylated human ANAPC1 (arrowhead). Lane 1 shows lysate from asynchronous cells. Lane 2 shows lysate from cells treated with nocodazole. While some phosphorylated ANAPC1 is present in untreated cell, the amount of phosphorylated protein is increased in cell preparations arrested in mitosis. Each lane contains approximately 30 ug of HeLa whole cell lysates, separated by 4-8% SDS-PAGE followed by transfer to nitrocellulose. After blocking the membrane was probed with the primary antibody diluted to 1 : 1,000 overnight at 4°C followed by washes and reaction with a 1 : 10,000 dilution of IRDye™800 conjugated Gt-a-Rabbit IgG [H&L] MX for 45 min at room temperature. IRDye™800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc.
Immunohistochemistry of ANAPC1 (phospho S377) polyclonal antibody (Cat # PAB9936) was used at 5.0 ug/mL to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows moderate positive cytoplasmic and occasional nuclear staining of pancreatic carcinoma cells at 60X. Tissue was formalin-fixed and paraffin embedded. The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain. Personal Communication, Tina Roush, Life Span Biosciences, Seattle, WA.
ANAPC1 is 1 of at least 10 subunits of the anaphase-promoting complex (APC), which functions at the metaphase-to-anaphase transition of the cell cycle and is regulated by spindle checkpoint proteins. The APC is an E3 ubiquitin ligase that targets cell cycle regulatory proteins for degradation by the proteasome, thereby allowing progression through the cell cycle.[supplied by OMIM