[Protein-analysis] Re: Western Blot Formalin Fixed Tissue

Dr Engelbert Buxbaum via proteins%40net.bio.net (by engelbert_buxbaum from hotmail.com)
Thu Nov 25 08:51:25 EST 2010


In article <mailman.35.1289798685.15153.proteins from net.bio.net>, 
r.zhang from auckland.ac.nz says...
> 
> I would like to know if it is possible to do western blotting experiment
> using formalin fixed frozen tissues of rat brain, thanks for that Rong

Difficult, as formalin cross-links proteins. The more common method is 
to rehydrate sections and elute proteins with hot RIPA-buffer containing 
detergents. The proteins are reduced, carboxymethylated and protease 
digested. The resulting peptides are separated by capillary 
electrophoresis and identified by MS/MS. For a review see Nirmalan et 
al.: Mol. Biosyst. 4 (2008) 712-20


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