Protein isolation from RNAlater stored tissue - (Jul/08/2008 )

Hi,

Has anyone done western and IP using protein isolated from tissue stored for a brief time (1 hour) in RNAlater (ambion)?

If not, do you see any theoretical problem with it?

Yours.

cellcounter

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Alright. As I generally do, I called up tech support and Yes, you can definitely do the Western.

http://www.ambion.com/techlib/tn/113/6.html

IP is not so sure, but there is at least one publication to it.

Pubmed ID - 11235903

Problem solved. Still, any dreadful stories of personal experience are most welcome

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Out of curiosity I googled RNAlater and this is what I found:

Isolating protein from RNAlater Solution-stored samples
Proteins are also preserved in RNAlater Solution. RNAlater
Solution will denature proteins; therefore, protein obtained
from samples stored in it will be suitable for applications such
as Western blotting or 2D gel electrophoresis, but not for
applications that require native protein.

Here is the link:
http://www.ambion.com/techlib/prot/bp_7020.pdf

So, I would probably rule it out for IP if I were you.

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Thanks smu. In fact the pubmed ID article they gave is about IHC, not IP, so it makes sense.

However, I will try out IP too, it all depends upon whether my Ab will recognize the denatured protein. As long as it does not crosslink proteins (which it doesn't), I can always try. Will let you know in this thread. Give me a week

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Plz, do and update us, and thx for sharing your experience!

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Western and IP both work as usual!

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