|H||Transfected Only||77||Rabbit IgG|
Western blot analysis of extracts from 293T cells mock transfected (-) or transfected with human DIXDC1 (hDIXDC1, +), using DIXDC1 (D7P6Q) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).Learn more about how we get our images
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 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.
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 June 2016
Protocol Id: 263
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.
DIXDC1 (D7P6Q) Rabbit mAb recognizes transfected levels of total DIXDC1 protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu579 of human DIXDC1 protein.
DIXDC1 (DIXIN, CCD1), is a Disheveled-Axin (DIX) domain protein that was first identified in zebrafish, where it was shown to positively regulate the canonical Wnt signaling pathway (1). In vitro studies subsequently showed that DIXDC1 activates Wnt signaling by binding to Disheveled (Dvl), resulting in the conversion of Dvl from a (latent) polymeric form to biologically active oligomers (2). In vivo studies in mice confirmed an important role for DIXDC1 in early development, by showing that DIXDC1 regulates both neural progenitor proliferation and neuronal migration, via Wnt-dependent and Wnt-independent mechanisms, respectively (3). Additional research suggests that DIXDC1 may function as a suppressor of metastasis by negatively regulating Snail1, implicating DIXDC1 in regulation of epithelial-mesenchymal transition (4). This function for DIXDC1 was shown to be dependent upon phosphorylation at Ser592 by MARK1 and MARK4 kinases (4).
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|14315S||100 µl (10 western blots)||$255.00.0|