how to isolate membrane protein from mammalian cells( t cells), and how to perform westren blotting to show this is expected protein. please any body suggest me the details protocol regarding this.
Thanks in advance
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#2
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Posted 21 February 2002 - 09:38 AM
Hi,
as first, you have to disrupt cells by sonication in an appropriate lysis buffer and second to separate plasma membrane on a sucrose gradient by ultra-centrifugation.
After this step, proteins are separated on PAGE for western blotting.
I send you by e-mail a PDF paper which describe this procedure for our protein of interest.
as first, you have to disrupt cells by sonication in an appropriate lysis buffer and second to separate plasma membrane on a sucrose gradient by ultra-centrifugation.
After this step, proteins are separated on PAGE for western blotting.
I send you by e-mail a PDF paper which describe this procedure for our protein of interest.
Bye.
#3
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Posted 17 November 2009 - 12:08 PM
scogne, on Feb 21 2002, 09:38 AM, said:
Hi,
as first, you have to disrupt cells by sonication in an appropriate lysis buffer and second to separate plasma membrane on a sucrose gradient by ultra-centrifugation.
After this step, proteins are separated on PAGE for western blotting.
I send you by e-mail a PDF paper which describe this procedure for our protein of interest.<p>Bye.
as first, you have to disrupt cells by sonication in an appropriate lysis buffer and second to separate plasma membrane on a sucrose gradient by ultra-centrifugation.
After this step, proteins are separated on PAGE for western blotting.
I send you by e-mail a PDF paper which describe this procedure for our protein of interest.<p>Bye.
can you sent me too.....i really need it thank u
#4
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Hmmm, I think it's working
Posted 17 November 2009 - 03:57 PM
You resurrected a post from 2002, so your chances of getting this protocol are small. Google search for subcellular fractionation protocols. There are a few good ones on the Nature protocols site too.
