rt-pcr data and western blot data not compatible - what could be the explanation (Sep/12/2006 )
I found several protein changed by 2D by comparing control and Durg treated group of gels.
Foutunatelu, I found several of them, I first verify them by 1D western blot, however, it seems that the 1D bands shows opposite direction of protein level change.
I use Real time PCR to verify the same protein on the gene expression level, however, there are also sevel gene changed in oposite direction.
Does anyone encounter such problem?
How can I interpret my result?
Thanks a lot.
I believe the result are true result, but the way to interpret is head cracking.
Often RNA levels and protein levels are regulated independently. Lots of microarray/proteomics comparisons demonstrate a low correlation between expression levels or changes between protein and RNA, so I wouldn't worry too much about the different results you are seeing. Actually, what you see is very interesting, and opens the door for you to think about biochemical regulation of these particular RNAs and proteins. I would be more concerned, however, if the 1D and 2D results were very different.
Yes, I will accept whatever result I have got.
Hope I can find out the mechanism of this phenomena.
Thank you for your suggestion.
The 1D and 2D result are not compatible in some of my target proteins.
I conclude that it could be the antibody recognize more than one kind of protein isoform, or because the inaccurate of the MS result.
If I change different brand of antibody to test, could the problem be solved?
Could there be any other explanation for it?
I think the ideas you have are reasonable possibilities. There's no way to be sure unless you try it!
That is what we call, re---------search.
I am very glad to dicuss with you guys.
Hope it works by changing antibody.
The human being is so complicated, beyond my intelligence a lot to understand it well.
there are so many possible areas of post transcriptional and translational regulation, its hard to know for sure... but these guys are right, its probably rarer to find a gene/protein with a tight mRNA/protein correlation than one without