Versatile toolbox for high throughput biochemical and functional studies with fluorescent fusion proteins.

ABSTRACT: Fluorescent fusion proteins are widely used to study protein localization and interaction dynamics in living cells. However, to fully characterize proteins and to understand their function it is crucial to determine biochemical characteristics such as enzymatic activity and binding specificity. Here we demonstrate an easy, reliable and versatile medium/high-throughput method to study biochemical and functional characteristics of fluorescent fusion proteins. Using a new system based on 96-well micro plates comprising an immobilized GFP-binding protein (GFP-mulitTrap), we performed fast and efficient one-step purification of different GFP- and YFP-fusion proteins from crude cell lysate. After immobilization we determined highly reproducible binding ratios of cellular expressed GFP-fusion pro

SUBMITTER: Pichler G 

PROVIDER: S-EPMC3350483 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature