In article <F5Au4z.890 at news.muni.cz>, "Vladimir Rotrekl" <rotrekl at chemi.muni.cz> wrote:
> Dear all,
> I have produced recently some mutant proteins, which are not really
> examples of protein stability. But they are at least able to stay in
> solution at 4C. The problem appeared when I tried to run native PAGE.
> Samples precipitated after addition of glycerol(20%) loading buffer.
> My qustion is if (and if yes then how) it is possible to use other
> compounds to prepare sample buffer for proteins (like in DNA 10% ficol or
> sucrose).
>> Thanks a lot
> Vladimir
Proteins that I have purified for sequencing and dont want to block the N-terminus I use a sucrose-based loading buffer described by BioRad. You can find it from them with instructions to use their PVDF membrane.
good luck,
peter
