IUBio

Lipids off from Protein-Homogenate?

Amanda Nouwens anouwens at RNA.BIO.MQ.EDU.AU
Wed Jan 27 16:32:06 EST 1999


To:             	protein-analysis at net.bio.net
From:           	"MURAT ERAVCI" <eravci at zedat.fu-berlin.de>
Subject:        	Lipids off from Protein-Homogenate?
Date sent:      	Wed, 27 Jan 1999 13:29:33 +0100

> I try to make 2D electrophoresis with rat brain tissue. Can somebody tell me
> how i can get the the lipids off from my Protein homogenate, without
> decreasing the amount of the proteins ?

> Thank You in advance
> 

Try a chloroform/methanol extraction (ratio 1:10 v/v).  Add this to your sample, 
store at -70 degrees celsius (minimum of 1 hour).  Centrifuge at ~14000g x 30 
min.  Remove supernatent and dry sample (use a speedvac).  Solubilise in 2D 
sample buffer as usual. 

Cheers, 

Amanda. 
 


---------------------------------------------------
Amanda Nouwens
Australian Proteome Analysis Facility (APAF)
Macquarie University
Sydney, NSW 2109
Tel: 02 9850 6209
Fax: 02 9850 6200
Email: anouwens at proteome.org.au



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