Western Blot

Susanne Kölare Jeffrey via methods%40net.bio.net (by sjeffrey from u.washington.edu)
Wed Oct 10 13:04:03 EST 2007


Dear Sudheendra,

When you perfusion fix a tissue through the injection of fixatives  
through the heart of the animal, you are basically preparing tissues  
for sectioning and immunohistochemistry. Fixatives (ex. 4%  
paraformaldehyde) crosslink proteins so that they cannot unfold. In  
western blotting, I have only used extracts prepared from fresh,  
unfixed tissue, since I want to  "strech" out/unfold proteins so that  
my antibodies will find their epitope(s). Many antibodies that has  
been made using (unfolded) synthetic peptides, only work in western  
and not in immunohistochemistry since in perfused or immersion fixed  
tissue the proteins are fixed in their native state.

Susanne

On Oct 10, 2007, at 4:09 AM, Sudheendra Rao N R wrote:

> Hello all,
> can we western blot using protein extract from the formalin fixed  
> tissues?
> Does perfusion of tissue (say brain) has anything to do with  
> western blot
> based protein detection?
>
> Thanks in advance
> Sudheendra.
> National Brain Research Centre
>
>
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