HELP! TBS for GST?

[Cornelius Krasel]

Michael DiDonato (michael.didonato at utoronto.ca) wrote:
> I would like to know if anyone has carried out a purification of a GST
> fusion protein using Tris-buffered saline instead of PBS as the lysis
> buffer?  If so, does the fusion still bind to the column using Tris? At
> what pH?

Well, I don't call it TBS but I routinely purify GST fusion proteins in
20 mM Tris, 200 mM NaCl, 2 mM DTT, 1 mM EDTA pH 7.6. Cells are lysed
in a little less NaCl (100 mM NaCl).

High pHs will denature GST. To clean my column I use 100 mM Tris 500 mM
NaCl pH 8.5.

--Cornelius.

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/* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */
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