[Protein-analysis] Analysis of small peptides: voltage & acrylamide

M O via proteins%40net.bio.net (by gawak_momo from msn.com)
Tue Mar 17 09:35:45 EST 2009


Hi everyone,

I am trying to separate 3 smalls peptides (15kDa, 9kDa and 7kDa) , I am using the technic of "tricine sds-page" by schagger H. et al. of Nature protocols
The difference between the 9kDa and 7 kDa is visible but I want to increase it, the problem is that the system of detection ( fluorescence) cannot work with gel of 16% of acrylamide so i am working on gel 10%, and i am going to see if i can go further in the percentage of acrylamide.But my problem is about the voltage that I apply to the gel:

On the net i can't find any informations about " how to determine the best voltage/current in a sds-page gel"
So how can I determine the best voltage/current to apply to my gel?

And in lab there's three types of acrylamide:

Acrylamide -Bisacrylamide 40% 19:1 Sigma 
Acrylamide -Bisacrylamide 40% 37.5:1 Merck
Acrylamide -Bisacrylamide 40% 19:1 Merck

I am using the last one, will I increase the resolution by using the 37.5:1? i don t know much about the difference between 19:1 & 37.5:1

Thanks for paying attention to this post 

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