[Protein-analysis] Re: heterooligomer or homooligomer

Nick Theodorakis nick_theodorakis at hotmail.com
Sat Nov 19 09:27:09 EST 2005


On Thu, 17 Nov 2005 17:59:02 -0500, SL <user at eamples.net> wrote:

>SL wrote:
>> Hi,
>> 
>>  From gel filtration + western, my protein looks much bigger (~ 200 kD) 
>> than its monomer size (~60 kD). I am just wondering if there is a method 
>> I can detect if it's a protein complex (with other proteins), or just a 
>> oligomer of itself.
>> 

>I mean to know this before I am starting to purify this complex.
>

It's going to be difficult to answer that question definitively
without further purification. If it's abundant enough, and you have an
antibody, you could something like an immuniprecipation followed by
examination of the pellet by PAGE (either 1D or 2D), but it will take
a lot of work to rule out non-specific binding.

The answer will most likely become clearer during purification, but
since you are going to purify it anyway, why is it so important to
know before you start? Arthur Kornberg once said, "don't waste clean
thinking on dirty enzymes."

Nick

-- 
Nick Theodorakis
nick_theodorakis at hotmail.com
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