Western blot analysis of extracts from various cell lines using QKI Antibody.
|REACTIVITY||H M R Mk|
|MW (kDa)||38, 42|
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
QKI Antibody recognizes endogenous levels of total QKI protein. This antibody detects QKI-5 and other isoforms.Species Reactivity:
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly202 of human QKI protein. Antibodies are purified by protein A and peptide affinity chromatography.
Quaking (QKI) is a member of the signal transduction and activator of RNA (STAR) protein family of RNA binding proteins (1,2). Mutations in the mouse Qki locus results in impaired myelin formation resulting in tremors (3). QKI proteins exist as homodimers, and disruption of the dimerization process is lethal in mice (2). QKI exists in different isoforms that differ in their C-terminus, resulting in unique subcellular localizations (4). The nuclear isoform of QKI, QKI-5, is involved in regulation of alternative splicing of MAG mRNA, which encodes for a protein important for myelin sheath formation and maintenance (4,5). QKI has also been implicated in schizophrenia and oligodendrocyte differentiation (6). QKI can function as a tumor suppressor, as it is regulated by p53 to stabilize miRNAs that regulate TGF-Β signaling (7).
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