Help with quantitative analyses of protein

Wolfgang Schechinger wolfsc at ibms.sinica.edu.tw
Thu Nov 15 04:02:57 EST 2001


Use high protein binding plates and probe with an AB or for function

Wo

> 
> 
> 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/
> >>
> >>
> > 
> > 
> 
> 


* * *
this is not a text message. 
it rather is an irrational number.
* * *
Dr. Wolfgang Schechinger
Institute of Biomedical Sciences
Academia Sinica, Taipei, Taiwan R.O.C.
* * *

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