Western blot analysis of extracts from various cell lines using DDX3 (D19B4) Rabbit mAb.
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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 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 June 2020
Protocol Id: 10
DDX3 (D19B4) Rabbit mAb recognizes endogenous levels of total DDX3 protein.
Human, Mouse, Rat, Monkey
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human DDX3 protein.
The DEAD box family of RNA helicases is characterized in part by a common D-E-A-D amino acid motif. The family is composed of a growing number of proteins found in a wide range of organisms from bacteria to mammals. DEAD helicases have distinct biological functions in RNA metabolism and ribonucleoprotein (RNP) processing (reviewed in 1,2).
DDX3 is a DEAD box family RNA helicase with diverse cellular functions. DDX3 is required for nuclear export of HIV-1 viral transcripts, possibly in a complex with the viral Rev protein and host cofactor CRM1 (3). DDX3 is required for hepatitis C virus (HCV) RNA replication (4) and its expression is downregulated in hepatitis B virus (HBV) associated hepatocellular carcinoma (HCC) (5).
Recent evidence suggests that DDX3 functions as a tumor suppressor protein. Its expression inhibits tumor cell colony formation and increases expression of the cdk inhibitor p21 Waf1/Cip1. Low DDX3 expression has been shown in HCC (5,6), and aberrant subcellular localization occurs in many squamous cell carcinomas (6). Reduced DDX3 expression in cultured cells causes a diminished dependence on serum for cell proliferation and changes in cyclin D1 and p21 Waf1/Cip1 expression (5).
DDX3 is phosphorylated at Thr204 and Thr323 by the mitotic cyclin dependent kinase, cyclin B/cdc2. This phosphorylation is thought to cause a loss of DDX3 function and a concomitant repression of ribosome biogenesis and translation in mitosis (7).
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