Hi Nick,
A couple of other suggestions:
(1) Try an E. coli K strain (e.g. JM109). They're different in
glucose metabolism (JN Phue et al, 2005, Biotechnol Bioeng 90:805820)
and in my experience tend to grow more slowly - which seems to help
improve yields of large constructs.
(2) Opt
Hello Anindito,
While Nick and Paul have thrown you some great ideas around the TAP-tag
theme (although GST may not be ideal as it dimerises itself), I feel that
I've read your initial e-mail differently from them. Maybe you could
clarify?
It sounds like you have a specific tag of ~5kDa in mind
Hi Careina,
Since alternative methods are being suggested...
ITC can be good for quantitating a monomer-dimer equilibrium by
diluting dimers out from a concentrated solution (which obviously
favours the dimer) - and presuming a reasonable Kon/Koff.
Alan Cooper has kindly figured out the data fi
Hello folks,
I would like to measure the concentrations of proteins containing
phosphotyrosines using absorbance at 280nm.
I'm wondering about calculating extinction coefficients by the Edelhoch
method - as used in ProtParam for example:
http://web.expasy.org/protparam/protparam-doc.html
Given