Urea on SDS-PAGE

[Bahara]

Hi everyone!

I intend to use an urea containing SDS-PAGE. My stacking and separating gels will contain urea, but do I have to add urea to sample buffer too?

Best!

[aftabac]

Hi

well in some recipies peoples do add urea in sample buffer, so you can add and check its effect.
you can run the gel in a way that in some sample buffer dont add urea and in other add urea and check the difference, then you can have better idea whether to use urea or not.

best luck......

regards
aftab

[deepee]

I totally agree with the previous suggestions. I regularly run samples that contain urea and those that don't I boil in SDS instead, this applies to normal SDS gels and those with urea (although it's been a while since I included urea in my gels). So no, it shouldn't be neccesary but you might get better results, it'd be worth trying both if you can.
deepee

[Bahara]

Thank you all.
Best!

[heidilwc]

hi all~i'm new to this forum as well as new to SDS-PAGE~
in fact , what's the use of adding urea to the gel and how will it improve the results?
thx for viewing~

[Bahara]

Hi

Well In "principles and reactions of protein extraction purification and characrtization" book we read : "SDS-Urea PAGE is often used for protein of low molecular weight and membrane proteins. In SDS-Page low molecular proteins may not be proportional to their molecular weight, as the protein charg properties become significant relative to the mass. SDS-Urea is sutible for membrane proteins, as they may not be soluble in at conditions used in SDS PAGE."

This book has helped me very much.

Best!