-----BEGIN PGP SIGNED MESSAGE----- Hash: SHA1 Dear Theresa,
you might also try changing the resin. I have worked on a protein which would no bind to Ni-NTA (Qiagen) at all but could greatly be purified when using Ni-IDA (then Pharmacia) instead. The protein expressed to about 60mg/l LB in E. coli and was well folded. Best wishes, Tim On 01/15/2012 07:23 PM, Theresa H. Hsu wrote: > Hi all > > I have a His-tagged soluble protein (8 His residues added to 90 kDa protein) > that do not bind to IMAC column based on flowthrough showing up with Western > blott. Do you have suggestions to improve the binding? > > Binding condition is 50 mM Tris-HCl 8.0, 300 mM NaCl, 10 mM imidazole pH to > 8.0. > > Thank you. > > Theresa > - -- - -- 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/ iD8DBQFPFASdUxlJ7aRr7hoRAgiUAKD6Rzwp4nEtT4R/2xycc+k4Z0oKqgCg1HOn HgglN/BkelWqK8AFbn+IYHg= =HrLG -----END PGP SIGNATURE-----
