Matt, there should be also a material called PBE94 for pH range 9 to 4
(and another one called Pharmalyte for high pH) to pack your own column.
It worked pretty well for our protein and is not in the high price
region of the MonoP column.
Best,
Guenter
Matthew Chu wrote:
Thanks Guenter and Andreas,
Yea, I have taken a look for the Mono P before, I thought the material
they used in Mono P is basically the same as in Mono Q and I found the
book "Protein Purification Protocols: Second Edition" by Paul Cutler
mentioned that phosphate buffer can also generate a continuous
gradient....that's why I reckon we can also perform pH gradient in
Mono Q. But I am worrying if it is a good idea to go for it, as both
mono Q or mono P are quite expensive.
Matt
2008/6/24 Andreas Förster <[EMAIL PROTECTED]
<mailto:[EMAIL PROTECTED]>>:
Hey Matt,
it seems to me that what you're asking for is chromatofocusing.
See the official GE documentation:
http://www6.gelifesciences.com/aptrix/upp00919.nsf/Content/WD:Chromatofocusin(260949098-R350)
<http://www6.gelifesciences.com/aptrix/upp00919.nsf/Content/WD:Chromatofocusin%28260949098-R350%29>
The proprietary buffers are a bit expensive, but as you found out,
they're a bit complicated to make. I don't know if you'd ruin
your Mono Q with a pH gradient. If in doubt, buy one of the
dedicated Mono P column.
Hope that helps.
Andreas
Matthew Chu wrote:
Dear All,
Sorry for off-topic question. Does anyone have any experience
in purifying protein using pH gradient in Mono Q column?
I have been googling for a whole day, only one paper was found
to mention performing pH gradient in Mono Q, but in a mixture
of amine buffering species, which is a bit too complicated (J.
Chromatogr. A 1164 (2007) 181 - 188. Can Tris-Cl/Tris-base or
phosphate buffer give a linear pH gradient from pH 8.0 to 4.0?
Is it usual to perform pH gradient in Mono Q as I don't want
to ruin my Mono Q column...
Any suggestions are welcome. Thanks in advance!
Kind regards,
Matt
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Matthew LH Chu
PhD Student
School of Pharmacy and Pharmaceutical Sciences
University of Manchester
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Matthew LH Chu
PhD Student
School of Pharmacy and Pharmaceutical Sciences
University of Manchester
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