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Anti-ANAPC1 PHOSPHO Antibody
Also for ANAPC1 (NM_022662)
|This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids 373-382 of Human Apc1 protein.|
||ELISA: 1:2,000 - 1:10,000, WB: 1:200 - 1:1,000, IHC: 5.0 ug/ml
|0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2|
|APC1 (also known as Anaphase promoting complex subunit 1, Cyclosome subunit 1, Protein Tsg24, Mitotic checkpoint regulator and ANAPC1) is 1 of at least 11 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.
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Western blot using Affinity Purified anti-APC1 pS377 antibody shows detection of a band ~215 kDa corresponding to phosphorylated human APC1 (arrowhead). Lane 1 shows lysate from asynchronous cells. Lane 2 shows lysate from cells treated with nocodazole. While some phosphorylated APC1 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 IRDye800 conjugated Gt-a-Rabbit IgG [H&L] MX (611-132-122) for 45 min at room temperature. IRDye800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.
affinity purified anti-APC1 pS377 antibody 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, LifeSpanBiosciences, Seattle, WA.