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sodium azide as protease inhibitor - protein overexpression (Aug/12/2005 )

Hi,
I came across a protocol where they add sodium azide to the E.coli culture before induction with IPTG in order to reduce proteolytic cleavage of an overexpressed protein. Has anyone tried this and does this work?

Thank you.

-yuliya-

I have never heard of this. Could you tell me what the reference for this is?

Daniel

DNA sequencing programs

-Daniel Tillett-

hi
sodium azide is an antibiotic usually used for preserving antibodies in TBS/PBS milk...
But as protease inhibitor? blink.gif
i'm interested in the reference too...
fred

BTW : sodium azide is now forbidden due to high toxicity.
For preserving solutions, i've saw about micr-o-protect...

-fred_33-

I thought that azide worked by decoupling the electron chain not as a protease inhibitor. I just looked up sodium azide in wikipedia and to quote "Azide anions prevent the cells of the body from using oxygen, inhibiting the function of cytochrome oxidase by binding irreversibly to the heme cofactor"

This doesn't sound like a protease inhibitor to me blink.gif

Daniel

Longer automated DNA sequencing reads

-Daniel Tillett-

I thought it was just a preservative. I too would be interested to know about it's protease inhibitor properties.
I do know that it causes problems when used to preserve antibodies labelled with HRP, but I'm not sure how.

-Rosie-