Western blot analysis of extracts from various cell lines using UAP56 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
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
UAP56 Antibody recognizes endogenous levels of total UAP56 protein.Species Reactivity:
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly24 of human UAP56 protein. Antibodies are purified by protein A and peptide affinity chromatography.
The UAP56 gene is found in the central MHC region and encodes a member of the DEAD-box family of RNA helicases (1). Also known as DDX39B and BAT1, UAP56 functions as an ATP-dependent splicing factor and RNA helicase in the evolutionary conserved transcription/export (TREX) complex. The TREX complex is recruited to sites of active transcription, where it travels along the length of the gene with RNA polymerase II and exports resulting mRNAs to the cytoplasm (2-8). Both UAP56 and its paralog DDX39A are hijacked by various viral replication machineries to enable viral reproduction and mRNA export (9-11). UAP56 and DDX39A have also been implicated in promoting the AR-V7 splice variant in advanced prostate cancers (12).
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