SDS-PAGE

Duncan Clark blackhole at abuse.plus.com
Mon Jul 17 11:25:25 EST 2006


Historians believe that in newspost <44bace96$1 at clarion.carno.net.au> on 
Mon, 17 Jul 2006, Bean Long <ben.long at yourfinger.anu.edu.au> penned the 
following literary masterpiece:
>> On 7/14/06, yan sun <yansun2005 at gmail.com> wrote:
>>> Hi all,
>>> I was trying to figure out if my expected protein was expressed. The
>>> molecular weight is about 32 KD. dimer.
>>> I run the SDS-PAGE, But I did not find the protein band around 
>>>33KD(on  the
>>> marker) [image: blink.gif]
>>> Basically, I just broke the cell through the ultrasonocation, and extract
>>> the protein, I did not do more purification.
>>> My question is: I did not find the protein band, is that because the 
>>>protein
>>> is too diluted in the solution?
>>> the second question: Since the protein is dimeric, should I try to 
>>>find the
>>> band around [image: blink.gif] 16KD? or both 32KD and 16KD?
>
>If you have an antibody then a western would clear this up.  As Pow 
>mentioned, you should be looking for the monomeric form (16 kDa). Was 
>the protein expressed strongly (e.g. with an inducible promoter)??  If 
>so, then you might see a strong band, otherwise it may be lost amongst 
>the other proteins in your mix.  Run a non-induced or wild-type protein 
>extract control alongside your expression sample to see if a particular 
>protein band has appeared.

If it is in inclusion bodies you may not have any soluble protein as it 
will be in the cell debris pellet.

Duncan
-- 
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.


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