Protogel vs biorad's a-bis acrylamide - better one? (May/11/2009 )

Hi!

I am trying to detect a small protein about 16kDa and am currently using biorad's a-bis acrylamide without much succes.. Somebody suggested me that Protogel apparently is better for small proteins.. I am not sure about this. Has anybody got any experience about this?

Thanks!

Use a higher percent gel or one with a low ratio of bis:acrylamide (e.g. 1:19)

bob1 on May 12 2009, 02:17 AM said:

hi

I have tried using both 12% and 15% gels. I can get bands using a control peptide (positive control) but not my samples

SO is it true that protogel is better than biorad's in this respect??

thanks

Are you talking about pre-cast gels? I pour my own, so I can't talk about the relative merits of different companies' pre-cast gels.

bob1 on May 13 2009, 02:34 AM said:

no I am not talking about pre-cast gels. I also make my own gels

have you tried the tris-tricine buffer system (shager and von jagow)?

it is better for separation of small proteins and peptides.

in fact, your 16kDa protein should separate nicely in a 10% gel with this buffer system.

you should be able to find it by searching the archives.

mdfenko on May 14 2009, 09:19 PM said:

Hi,

Thanks for the reply, yes I have tried using Tricine buffer, but with 12% gel instead. I haven't tried using 10% for that, wouldn't it be too large(pore size), for my small protein? Also, do you know if protogel is better for small proteins? I could not find any literature regarding the same

cupidstunt on May 15 2009, 06:24 AM said:

it's not so much the pore size as the trailing ion that has the desired effect with tricine and acrylamide concentration of 10% or more. by the way, what was the result with tricine and 12% gel?

i'm not familiar with protogel so i can't properly respond to that part of your question.