Hi,
I had two samples (with or without treatment) sent for untargeted proteomics analysis through mass spectrometry.
We found there is a change in amount of an enzyme.
I want to know if I need to use western blot to confirm this change by western blot? as western blot is not very sensitive and as accurate by LC-MS.
Hi there,
What you actually need is an internal standard to normalize the results.
What if you go for SRM for the enzyme, since you have the library info, this would quantify easily.
If you are thinking wether you need to confirm a relative change in concentration observed between these two samples that is observed in your mass spectrometry data by another analysis method then I would say no.
An absolute quantitation would require a new type of measurement though, and I agree with Akita that a quantitative SRM mass spec is your best bet.
If you want to confirm the results of this one against one sample experiment however then you need to generate some statistics for that. And that ideally will require some form of power calculation involving both the observed difference and the level of confidence you want to have in your end result.
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