Protocol

Abbreviation

NU_default_purification

Name

NU tag-on purification

Laboratory name

Northwestern University

Type

purification

Description

Proteins are purified by Ni-affinity chromatography on HiTrep FF and desalting HiPrep 20/10 columns using AKTA Express system. Pure proteins are collected in buffer, containing 10 mM Tris-HCl (pH 8.3, 500 mM NaCl and 5 mM b-mercaptoethanol. Samples are concentrated to 5-25 mg/ml depending on solubility and used for crystallization experiments. Unused protein is flash-frozen in liquid nitrogen and stored at -80C.