Membrane protein isolation??

Dima Klenchin klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu
Fri Mar 12 21:21:39 EST 1999


In article <7cc2al$b7m at ds2.acs.ucalgary.ca>, Neal Melvin <nrmelvin at acs.ucalgary.ca> wrote:
>This is a multi-part message in MIME format.
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Could you please not post MIME stuff, and use plain text instead?

>Can anyone recommend a good starting protocol for the isolation of
>membrane proteins from molluscan (or any neurons to begin with) neurons?

All protein purification is alike. That is, it is very different, highly 
empirical, and unpredictable.  Unlike nucleic acids, there are no 
ready-to-use recipies. Almost everyhting depends on your protein and
application. The list of things to ask that helps to sort out countless 
possibilities is HUGE. Please provide as much background info as
possible, only then a reasonable advice can be tried. Partial list in no 
particular order:

Amount of source material, abundance of protein, method of detecion, 
aim of puirification, equipment available, MW of the protein, 
biological activity, homologes in other tissues/species, etc, etc. 

Without additional info, the only advice possible sounds as fiollows:
Solubilize membranes (well-washed under stringent conditions that 
still keep the protein insoluble) in cheapest detergent that preserves
desired properties of your protein and apply usual fractionation 
techniques such as precipitation, various column chromatographies, and 
electophoresis to purify your protein. - See how pointless it is?

- Dima



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