|H M R Mk||Endogenous||70||Rabbit|
Western blot analysis of extracts from various cell lines using PRC1 Antibody.Learn more about how we get our images
Western blot analysis of extracts from HeLa cells, untreated (-) or nocodazole-treated (+; 100 ng/ml overnight) to induce mitotic arrest and increase CDK1 activity. Increased CDK1 activity induces PRC1 phosphorylation, as indicated by the upper band.Learn more about how we get our images
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised October 2016
Protocol Id: 10
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
PRC1 Antibody recognizes endogenous levels of total PRC1 protein.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly522 of human PRC1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Protein Regulator of Cytokinesis 1 (PRC1) is a member of the MAP65/ASE1 family of nonmotor microtubule-associated proteins, first described in budding yeast (1,2). PRC1 is a substrate of CDK1, which maintains PRC1 in an inactive, monomeric state (2). Cell-cycle dependent degradation of CDK1 leads to dephosphorylation of PRC1 and subsequent KIF4-mediated translocation to the plus ends of microtubules, where it promotes microtubule bundling by cross-linking antiparallel microtubules (3-5). The microtubule bundling functions of PRC1 play a critical role in maintaining structural integritry of the spindle midzone during cytokinesis (3,6,7).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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