Western blot analysis of lysates from various cell lines using CK2α Antibody.
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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.
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
This antibody detects endogenous levels of total CK2α 1 protein. This antibody may cross-react with CK2α prime.Species Reactivity:
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acid residues near the carboxy-terminus of human CK2α1. Antibodies are purified by protein A and peptide affinity chromatography.
CK2 (formerly called Casein Kinase II) is a highly conserved protein kinase with more than 300 substrates regulating cell growth, cell death, and cell survival. CK2 has been implicated in the response to UV irradiation-induced DNA damage, targeting XRCC1 (1) and BRCA1 (2) as well as regulating p53 tumor suppressor protein functions (3). Furthermore, CK2 plays a key role in NF-κB activation (4). UV irradiation stimulates CK2-mediated phosphorylation of several carboxy-terminal residues within IκBα, resulting in IκBα proteasomal degradation and the release and nuclear translocation of active NF-κB. CK2 is also dysregulated in many cancers (5) and neurodegenerative diseases such as Alzheimer's and Parkinson's diseases (6). Structurally, CK2 is a multimeric protein complex consisting of two catalytic subunits (α or α') and two regulatory β subunits (7). CK2 is distributed ubiquitously and is apparently constitutively active (7). While cell cycle-dependent Ser-Pro phosphorylation sites have been identified on CK2α and CK2β, Tyr255 phosphorylation by the Src-related kinase c-Fgr seems to have the greatest effect on CK2α activity (8,9).
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