Friday, July 31, 2009

Western blotting with an anti-HuR monoclonal

The HuR protein is an important regulator of gene expression. It has been shown in many studies that HuR mediates the effects of cellular signals that upregulate gene expression at theRNA level. In general, such signaling pathways lead to an increased binding of HuR to particular elements within the mRNA, which abrogates the effect of factors that degrade mRNA. More recently, HuR has been identified as an important cofactor in the action of microRNAs. Clonegene has developed high affinity monoclonal antibodies to HuR that have been used in many of these studies. CG001 works well in western blot analysis. CG001 is supplied as a purified monoclonal. For westerns, use at 1/1000 ( 2 hours incubation at RT ) or 1/5000 ( overnight in cold room) and a standard anti-mouse secondary coupled to ECL detection. The figure show the reactivity of CG001 with extracts ( 50ug protein per lane ) from Hela cells. Lane 1 is no antibody , lane 2 is CG001 , lane 3 CG001 competed with antigen peptide , lane 4 CG001 competed with irrelevant peptide and lane 5 irrelevant antibody. The apparent molecular weight of HuR is 36 kD.
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