Peptide fixation

InYup Park ipark at bioneer.kaist.ac.kr
Fri Sep 26 21:04:40 EST 1997


Betsy Lytle wrote:
>
> I forgot to mention in my post yesterday that I am using Tricine gels.
>
> >Hi all,
> >
> >I am analyzing a 3.5 kD protein (pI=9.3) by SDS-PAGE and am having problems
> >visualizing it. This could be due either to poor staining or to diffusion
> >of the protein out of the gel. Staining by Coomassie blue and silver stain
> >are both poor, which makes me think diffusion is the problem. I have tried
> >fixing the gel with 12% TCA/3.5% sulfosalicylic acid without success. I may
> >try using glutaraldehyde next, but would like any advice from others who
> >have had this problem.
> >
> >Thanks,
> >
> >Betsy Lytle

Have you checked the pH of your buffers? (gel buffer and the running
buffers)

Also, are you using a stacking gel?  If so, you may need to adjust its
length.

Hope this helps.


InYup Park



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