Help with quantitative analyses of protein

Trond Erik Vee Aune teva at online.no
Thu Nov 15 03:51:54 EST 2001



Silas Bruun wrote:

> What about setting up an ELISA reaction. That can be made quantitatively and
> there will not be any problem with a smear - as in your WB


Is is easy to do ELISA on inclusion bodies? Wound't it be difficult to 
bind the denaturated protein we get from the inclusion bodies to the 
well? All the ELISA I've performed have been based upon the fact that 
you bind the protein of interest to the well via an antibody that 
recognizes the protein based on it's folding, a folding a denaturated 
protein probably lacks.

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


> 
> cheers
> silas
> 
> --
> 
> 
> 
> ***************
> Silas Bruun
> The Protein Laboratory
> Institute of Molecular Pathology
> University of Copenhagen
> Blegdamsvej 3 Bldg. 6.2
> 2200 Cph N, Denmark
> fax: +45 353-60116
> "Trond Erik Vee Aune" <teva at online.no> wrote in message
> news:3BF28067.3070006 at online.no...
> 
>>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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