viewing GFP protein on Western blot

Frank O. Fackelmayer fof1 at chclu.chemie.uni-konstanz.de
Wed Aug 13 03:41:53 EST 1997


In article <5sql3q$iol$2 at nntp.ucs.ubc.ca>, lpss at unixg.ubc.ca (Alex Chang) wrote:

> Alex Chang (lpss at unixg.ubc.ca) wrote:
> : I am planning to do immunoprecipitation of my GFP-tagged protein, after 
> : SDS-PAGE and transblotting onto a membrane, look at the GFP-tagged protein 
> : directly by a UV light source without using antibody detection (or simply 
> : look at the gel directly without transblotting).
> 
> : I'd like to know anyone has done this, and what kind of UV source can be 
> : used.
> 
> 
> : Alex Chang
> : Pathology
> : University of British Columbia
> : achang at hivnet.ubc.ca
> 
> --
> Alex Chang
> Pathology
> University of British Columbia
> achang at hivnet.ubc.ca


Hi Alex,
I haven´t done it, and so have no evidence that I´m right, but I expect
that it won´t work. That´s because GFP-fluorescence is very sensitive to
denaturation. Actually this is one big disadvantage of GFP, because even
fixation of cells by organic solvents or formaldehyde kills the
fluorescence (as reported several times in the newgroups). So,
denaturation by SDS is likely to do the same. Please tell me if I´m wrong,
it would be so nice to visualize GFP in this way...

Sorry that I can´t help more,
Frank



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