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That's why I like HIS-SELECT a lot - low Imidazole elution (typically around 50 mM) and low nonspecific binding. Sigma charges a lot for it though. Artem -----Original Message----- From: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of Martin Hallberg Sent: Sunday, October 08, 2006 5:43 PM To: Nat Echols Cc: ccp4bb@dl.ac.uk Subject: Re: [ccp4bb]: off-topic: Ni purification media *** For details on how to be removed from this list visit the *** *** CCP4 home page http://www.ccp4.ac.uk *** 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
