-----BEGIN PGP SIGNED MESSAGE----- Hash: SHA1 Dear Sangeetha,
provided you express the protein in E. coli, you could also sonicate the cell debris in various buffers and compare supernatant/pellet on SDS-gel. It might already give you a first clue and is faster, cheaper and you don't risk to clog the new columns with aggregated protein. Tim On 02/27/2012 05:01 PM, Sangeetha Vedula wrote: > Dear bb users, > > I am trying to crystallize a ~320 kDa protein that crashes out if > concentrated past about 3 mg/mL. > > I would like to try to exchange it into various buffer-salt-additive > combinations to see which buffer works. For a starting point, I'd like to > use desalting colums. > > Does anyone have suggestions for good buffer exchange and sample recovery? > I woud like to load about 250 uL onto each column. > > Thanks a lot! > > Best regards, > > Sangeetha. > - -- - -- Dr Tim Gruene Institut fuer anorganische Chemie Tammannstr. 4 D-37077 Goettingen GPG Key ID = A46BEE1A -----BEGIN PGP SIGNATURE----- Version: GnuPG v1.4.10 (GNU/Linux) Comment: Using GnuPG with Mozilla - http://enigmail.mozdev.org/ iD8DBQFPS6xEUxlJ7aRr7hoRAqOqAJ0cYn4488ctcIRUeZa4K3f2smojfQCeKBDI +nO8deQmkPNB9nWSWJtBoOI= =SU1z -----END PGP SIGNATURE-----
