|H Mk||Endogenous||175||Rabbit IgG|
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, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
PICH (D4G8) Rabbit mAb recognizes endogenous levels of total PICH protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly610 of human PICH protein.
PICH is a helicase of the SNF2 family of ATPases and is essential for proper chromosome segregation during mitosis (1). While PICH was originally proposed to participate in spindle assembly checkpoint signaling (1), that function was subsequently called into question (2). When phosphorylated at Thr1063 by CDK1, PICH binds the polo-box domain of the mitotic kinase PLK1 (1) and targets it to chromosome arms (3), where it appears to facilitate proper chromosome arm cohesion (4). PICH is also a substrate of PLK1 (1). Localized to the cytoplasm during interphase, PICH begins to accumulate at centromeres and kinetochores in prometaphase (4). As chromosomes begin to separate at the onset of anaphase, PICH associates with ultrafine threads between sister centromeres thought to be composed of entangled DNA (5), a natural consequence of DNA replication. PICH is proposed to cooperate with BLM, a RecQ-like helicase implicated in the genetic disorder Bloom’s Syndrome, to displace centromeric histones along these threads, thus enabling them to span large distances without breaking (6). This provides a temporal window for topoisomerase IIα-mediated disentanglement (7). Defects in PICH or BLM disrupt proper chromatid segregation and result in the formation of micronuclei (6).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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|8886S||100 µl||$ 255.0|