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On Oct 8, 2006, at 10:46 PM, Nat Echols wrote:
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?
Same but different ;-) The possibility to elute at low imidazole
concentration (even lower than Talon) may be appreciated by your
protein.
Check out their protocol at:
https://www.macherey-nagel.com/web/MN-WEB-Biokatalog.nsf/web/
PROTINOTED/$File/ProtinoNi_TED_R03.pdf
Also, which Sepharose can take 1MPa? I haven't found anything
capable of more than 0.3, even the "Fast Flow" columns.
None, according to Amersham-Pharmacia-GE Healthcare. As Artem said in
his previous posting, you can often use a pressure
up to 1 MPa with Fast-flow resin (your mileage may vary.. depending
on a lot of factors..).. Avoid the cheap HiTrap columns for
these experiments.
Cheers,
Martin
PS. I have no connection with Macherey-Nagel (more than that they
gave me 5 g Ni-TED resin for free to try out...)
PS2. They also sell a more traditional IDA-based resin. I haven't
tried that but note that it has the same pressure
limits as the Ni-TED.
.
B. Martin Hallberg, PhD
Structural Genomics Consortium
Department of Medical Biochemistry and Biophysics
Karolinska Institutet
Scheeles väg 2
SE-171 77 Stockholm
http://sgc.ki.se/groupSB2.html
