SDS-PAGE with small proteins

Pow Joshi via methods%40net.bio.net (by pow.joshi At gmail.com)
Mon Nov 27 19:25:51 EST 2006


On 11/27/06, Marcin <marcin.j.wozniak At manchester.ac.uk> wrote:
> Hi everybody,
> I wonder if anyone knows how to separate and visualise small proteins
> (4-10 kDa) on a gel.
First, would you like to say what are the protein sizes you are looking for?

 I tried gradients, high percentage gels and
> Tris-tricine gels w/o any luck.
Tris-tricine gels should work well. Which company "assembly" are you
using? what percentages are you using? I have used 15% regular tris
glycine gels to resolve upto 3kDa.
 When I stain with Coomassie or Ponceau
> (after transfer onto 0.2 u nitrocellulose) I can see nice bands up from
> 10 kDa. Below that point nothing shows up! Also, I tried blotting with
> specific abs w/o any success. I would appreciate any suggestions.
What sort of protein concentrations are you expecting for the "small
proteins"? Having positive controls alongside helps to know where the
problem could be?

we could help you troubleshoot better if you gave more details.

best
pow

> Cheers, Marcin




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