[3dem] protein localization

[rkhayat at ccny.cuny.edu]

Hi,

I may be suggesting something you've already tried, but why not try some of the traditional membrane protein proteolysis experiments? 
     1. Proteolyse the protein with a protease to make sure it can be digested
     2. Decorate the liposome with the protein
     3. Purify the protein decorated liposome from free protien
     4. Treat the decorated liposome with protease 
     5. Test to see if the protease can digest in the presence of liposomes by running the same proteolysis experiment in the presence of the liposomes and a digestible protein that is not incorporated into the liposome

Reza Khayat, PhD
Assistant Professor
The City College of New York
160 Convent Avenue
Department of Chemistry
MR-1335
Tel. (212) 650-6070


---- Original message ----
>Date: Fri, 8 Feb 2013 15:56:06 +0000
>From: 3dem-bounces at ncmir.ucsd.edu (on behalf of Kimberley Cheng <Kimberley.Cheng at ki.se>)
>Subject: [3dem] protein localization  
>To: "3dem at ncmir.ucsd.edu" <3dem at ncmir.ucsd.edu>
>
>   Hi,
>
>   My colleague is working with protein-liposome
>   complexes. She thinks the proteins are either
>   incorporated in the liposomal membrane, or attached
>   to the surface of the liposomes. Is there any EM
>   method, or some other technique, that can be used to
>   assess where the proteins are located? Someone
>   suggested immunogold-labelling. Any thought about
>   this?
>
>   All the best,
>
>   Kimberley
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