*** For details on how to be removed from this list visit the *** *** CCP4 home page http://www.ccp4.ac.uk ***
Highly crosslinked sepharose has decent flow rates and decent pressure characteristics. I routinely run these types of columns at 1 MPa even though manufacturers typically recommend 0.4-0.6 MPa maximum. I believe that Poros media has much higher pressure limit. High-pressure columns tend to be more expensive than lower-pressure ones, of course.
Yikes. The stuff we get from Amersham has pressure limits around 0.3MPa (if that), which is very easy to exceed. (It's lower for gel filtration, but we're used to that.) We have multiple FPLCs - the problem now is I can't really take advantage of them for this step of purification. Part of this is a limited selection of column hardware; the gravity columns I use are significantly wider than any of the casings for FPLC use. If I'm going to spend >$500 on a new casing so I can use higher pressure, I might as well get the pricey media too.
The silica-based Ni-TED resin sold by Macherey-Nagel can take 10MPa pressure (~1400 psi for the SI-challenged) while the Poros MC resin takes 17 MPa (~2400 psi). Last time I checked, Ni-TED resin was sold at about the same price as the 1 MPa Pharmacia Sepharose (Swedish prices). Make sure that all parts of your purification setup can take these pressures.
I'll look into that media for sure - does it work well? Also, which Sepharose can take 1MPa? I haven't found anything capable of more than 0.3, even the "Fast Flow" columns.
thanks, Nat
