some
peptide bonds. Try His tag, then you don't need to use proteases for
cleavage, you can use Ni column.
Maia
- Original Message -
From: "Ashok Ranjan Nayak"
To:
Sent: Sunday, August 29, 2010 5:28 AM
Subject: [ccp4bb] Query regarding GST fusion protein purification
It sounds like list your GST construct is not binding to the column
(or very well) when the peptidase is attached. GST needs to form a dimer
to binding to the column - I suspect that your construct interferes with
dimer formation - when the peptidase is present, but when not there due
to stall
Hello one and all !!
I have been working on cloning and purification aspects on a Leishmanial
cysteine peptidase (pI= 8.2) for quite some time. I had cloned it in pGEX
expression vector. Expression seemed quite okay when induced with 0.5 and 1mM
IPTG, so did the solubility in 4 buffers at diffe
