IP Protocols and hints

AK via methods%40net.bio.net (by akhan357 from sbcglobal.net)
Thu May 22 15:46:28 EST 2008


I recently succeeded in endogenous co-IP of some erythrocyte 
membrane/peripheral proteins. the protocols and recipes in paper below may 
provide you with beginning at least. as you know IP is trial and error with 
different buffers/ detergents/ etc. However, you mention that you want to 
purify?? this protein by IP? IMHO you may end up wasting a lot of time for 
unusable amount of protein.
AK

J Biol Chem. 2008 May 23;283(21):14600-9. Epub 2008 Mar 17.
Dematin and Adducin Provide a Novel Link between the Spectrin Cytoskeleton 
and Human Erythrocyte Membrane by Directly Interacting with Glucose 
Transporter-1.


"Bean Long" <ben.long from yourfinger.anu.edu.au> wrote in message 
news:4830d6d8$1 from clarion.carno.net.au...
> Hi all,
>
> I've recently got my hands on an antibody to a small protein that's part 
> of an enormous complex (over 100 nm in diameter) and thought I might try 
> some IP to purify it. I've also got some Protein G-sepharose beads in the 
> fridge which I could use for IP. Can anyone offer protocols or point me in 
> the direction of some to help me out? I'll need to do this under native 
> conditions. I've done very little IP before, so any hints would be handy.
>
> Cheers,
>
> Bean 




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