The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/100 - 1/250.
1/500. Detects a band of approximately 72 kDa (predicted molecular weight: 72 kDa).
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Application notesIs unsuitable for Flow Cyt or IP.
FunctionCross-links antiparrallel microtubules at an average distance of 35 nM. Essential for controlling the spatiotemporal formation of the midzone and successful cytokinesis. Required for KIF14 localization to the central spindle and midbody.
Sequence similaritiesBelongs to the MAP65/ASE1 family.
DomainMicrotubule binding occurs via a basic patch in the central spectrin-like domain and requires also the unstructured C-terminal domain.
Post-translational modificationsPhosphorylation by CDK1 in early mitosis holds PRC1 in an inactive monomeric state, during the metaphase to anaphase transition, PRC1 is dephosphorylated, promoting interaction with KIF4A, which then translocates PRC1 along mitotic spindles to the plus ends of antiparallel interdigitating microtubules. Dephosphorylation also promotes MT-bundling activity by allowing dimerization.
Cellular localizationNucleus. Cytoplasm. Cytoplasm > cytoskeleton > spindle pole. Predominantly localized to the nucleus of interphase cells. During mitosis becomes associated with the mitotic spindle poles and localizes with the cell midbody during cytokinesis.
Dot Blot - Anti-PRC1 (phospho T481) antibody [EP1514Y] (ab62366)
Primary ab dilution 1:1000 dilution. Secondary ab description and code (ab id) Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051). Secondary ab dilution 1:100,000 dilution. Blocking buffer and concentration 5% NFDM/TBST. Lane 1: PRC1 (pT481) phospho peptide Lane 2: PRC1 non-phospho peptide. Exposure time 3 minutes
Western blot - Anti-PRC1 (phospho T481) antibody [EP1514Y] (ab62366)
All lanes : Anti-PRC1 (phospho T481) antibody [EP1514Y] (ab62366) at 1/500 dilution
Lane 1 : HeLa cell lysates (untreated) Lane 2 : HeLa cell lysates treated with aphidicolin + nocodazole
Lysates/proteins at 10 µg per lane.
Secondary goat anti-rabbit HRP labeled, at 1/2000 dilution
Predicted band size : 72 kDa Observed band size : 72 kDa
References for Anti-PRC1 (phospho T481) antibody [EP1514Y] (ab62366)
This product has been referenced in:
Cundell MJ et al. A PP2A-B55 recognition signal controls substrate dephosphorylation kinetics during mitotic exit. J Cell Biol214:539-54 (2016).
Read more (PubMed: 27551054) »