mRNA level and protein level correlation?

Dr Engelbert Buxbaum engelbert_buxbaum at hotmail.com
Mon Mar 22 08:27:40 EST 2004


"Michael L. Sullivan" wrote:

> I'd 
> be inclined to believe the rtPCR results, but depending on just how 
> you did the experiment, your western blot numbers my be suspect.  

Quantitative PCR is a much bigger kettle of fish than many people think.
There were a couple of papers in the last BioSpektrum on that issue. 

> I 
> think it is very hard to get quantitative data from a western both 
> because of the high degree of variation one gets within a single 
> blot, and also because western signals are very, very non-linear.  

Dot blots evaluated on a 96-well type chemoluminescence reader are much
more repeatable, and are linear over 4-6 orders of magnitude. Of course
you need to verify in western that there is only one protein interacting
with the antibody. 





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