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Topic: Insert immunoprecipitated proteins into micelle  (Read 2196 times)

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Offline Abey303

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Insert immunoprecipitated proteins into micelle
« on: July 20, 2013, 02:47:01 PM »
Hi all,

I'm wondering if anyone knows a way to insert immunoprecipitated proteins from cellular lysates into micelle so that the proteins can be stuck on the lipid?

Thanks in advance.

Abey


Offline Arkcon

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Re: Insert immunoprecipitated proteins into micelle
« Reply #1 on: July 20, 2013, 03:22:40 PM »
Hmmm ... tricky.  OK, so you're building lipid micelles, presumably by sonicating them.  And you want externally bound lipophillic proteins -- not transmembrane proteins.  Is there any reason why simple mixing doesn't work?  (As in, did you try it and have it not work?)  Are you sure your protein immuno-complex is lipophillic enough?  Why, exactly, do you need micelles with immuno-complexs on them, anyway?  I mean, I'm sure you have a reason, I'm just curious as to what the application is.  Did you read of this procedure in a peer-reviewed journal?  But then, that's where the protocol ought to be.
Hey, I'm not judging.  I just like to shoot straight.  I'm a man of science.

Offline Abey303

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Re: Insert immunoprecipitated proteins into micelle
« Reply #2 on: July 22, 2013, 02:43:41 PM »
Hi Arkcon,

Thank you so much for your reply.
I'm reallly not familiar with lipid biology so probably I didn't express my question well.
The protein we're interested in is indeed a transmembrane protein. We usually lyze the cells in NP40 lysis buffer and immunoprecipitate our protein of interest. However, in this case, the proteins would be all released from the membrane. So I guess my question is do you know a way to lyze the cells while keeping the transmembrane proteins still stuck in the membrane? And can I do Ip after that?

Thanks a lot!
Abey

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