3 questions

Brian P Higgins via methods%40net.bio.net (by higginsb78 from gmail.com)
Fri May 23 13:00:30 EST 2008


Regarding point 3, you are comparing two methods that differ greatly in
their sensitivity.  I wouldn't expect them to match up to the same degree.
You also need to take into account whatever you are using to
standardize/normalize in each case.  Assuming a linear relationship and
perfect correlation between mRNA levels and protein can be dangerous.

On Fri, May 23, 2008 at 6:17 AM, Kamalian, Laleh <L.Kamalian from liverpool.ac.uk>
wrote:

> I have 3 questions if someone could help me with those please.
> 1. Could 2 different bands (size wise) be detected for the same protein
> using poly colonal and monoclonal antibodies?
> 2.  If the samples are run on electrophoresis for too long, can the size of
> the protein show difference comparing to the previous (shorter) runs?
> 3.  In siRNA , I have knock down my gene of interest at the mRNA level
> (70-80% according to quantitative PCR x2 experiments), but western blot of
> the same colony cell protein lysates show no or less ( only 30% ) decrement.
>  Any explanation please?
>
> Thanks
> Laleh
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