protein problems on gels

Richard Van Frank vanfrank at iquest.net
Sat Jan 4 12:55:38 EST 1997


In article <32CED947.24E1 at jcu.edu.au>,
   lachlan harris <lachlan.harris at jcu.edu.au> wrote:
>Can anyone help me with this problem? I have a pure microbial protein 
>which when run on an SDS non-reduced (ie no mercaptoethanol) PAGE gel 
>runs at approximately 50 KDa. When the same protein is run under 
>SDS-reduced conditions two subunits appear- one running at approx 40 KDa 
>and the other at approx 60 KDa.
>
>It was my understanding that the two subunits should add up to be the 
>same size as the protein run under SDS-non reduced conditons. But this 
>isn't happening. Does anyone know why?
I don't believe this is a subunit problem. The subunit can't be larger than the 
original protein. I believe you are not geting a complete reduction but not 
knowing the conditons you are using I can't say for sure. Try more sever 
reduction conditions. 




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