Help with quantitative analyses of protein

Trond Erik Vee Aune teva at online.no
Wed Nov 14 09:32:07 EST 2001


Hi all!

How do you measure the amount of protein from expression studies? We've 
been trying to quantify the yield from protein gels and from Western 
blots, but without much success. The protein is denaturated from 
inclusion bodies, and what we see is more of a smear than distinct 
bands. So perhaps our method for preparing the samples are flawed. Do 
you know of any good protocols for preparing pellet fractions containing 
inclusion bodies that shall be run on gels and blotted? Does it happen 
that not all of the inclusion bodies are avaiable for antibody binding 
and hence detection?

The software is not helping either. We currently use Bio-Rads Quantity 
One software, do you know of any other software that may do the trick, 
that is to quantify bands from either gel or blot and estimate yield 
compared to a standard?

Kind regards,
Trond Erik Vee Aune
http://kibt.chembio.ntnu.no/molgen/




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