The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/10000. 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.
1/100 - 1/250.
1/20. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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.
Immunofluorescent staining of HeLa cells using ab51248 (1:100).
Immunocytochemistry/ Immunofluorescence - PRC1 antibody [EP1513Y] (ab51248)This image is courtesy of an anonymous Abreview
ab51248 staining PRC1 in human breast cancer cells by ICC/IF. The cells were paraformaldehyde fixed and blocked in 1% serum for 1 hour at 37°C without permeation step. The primary antibody was diluted 1/100 (PBS) and incubated with sample for 1 hour at 20°C. An Alexa Fluor® 488 conjugated donkey polyclonal to rabbit IgG, diluted 1/200 was used as secondary.
Overlay histogram showing HeLa cells stained with ab51248 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab51248, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
References for Anti-PRC1 antibody [EP1513Y] (ab51248)
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