Western blot analysis of extracts from various cell lines using CDK10 (D8Z7Z) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human CDK10 (hCDK10-Myc/DDK; +), using CDK10 (D8Z7Z) Rabbit mAb (upper) or Myc-Tag (71D10) Rabbit mAb #2278 (lower).
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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.
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
CDK10 (D8Z7Z) Rabbit mAb recognizes endogenous levels of total CDK10 protein. This antibody is not expected to react with CDK10 N-terminal deletion isoforms.Species Reactivity:
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val24 of human CDK10 protein.
Cyclin Dependent Kinase 10 (CDK10) is a Cdc2-related protein kinase that binds to and inhibits the transactivation activity of the transcription factor Ets2 (1).
CDK10 is activated by cyclin M, which is mutated and unable to activate CDK10 in the human developmental disorder, STAR syndrome. Phosphorylation of Ets2 by CDK10/Cyclin M leads to degradation of Ets2 by the proteasome (2). CDK10 also plays a role in the development of the zebrafish nervous system (3). Studies have shown that expression of CDK10, which is modulated by promoter hypermethylation, is decreased in human cancer (4-6). Further, studies show that CDK10 expression in breast cancer affects response to tamoxifen (7), and is correlated with disease progression (8). CDK10 regulates the expression of c-RAF, and signaling through the MAPK pathway (2-3, 6-7).
Explore pathways + proteins related to this product.
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