EDTA in lysis buffer?

Michael J. Morales mmorales at acpub.duke.edu
Wed Nov 5 12:06:22 EST 1997


user at mpi.com wrote:

> I've been playing around with conditions for lysis of
> mammalian cells
> lately (for subsequent IP or western analysis), and I'm
> wondering about
> the advantages and disadvantages of including EDTA.  Many
> people do; many
> others don't.  I've heard some stories of protein
> interactions breaking up
> as a result of the chelating properties of EDTA, which I
> suppose could be
> a potential disadvantage if studying specific
> interactions.  What are the
> advantages?  Inhibition of enzymatic activities (like
> proteases)?
> Cleaning up non-specific interactions?  Any other
> thoughts?
>
> -HS

Addition of EDTA inhibits metaloproteases, so is usually
desirable.
However, in some cases, such as downstream use of an
immobilized
metal column, EDTA must be avoided.

Hope this helps

Mike Morales





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