Optimization of formaldehyde cross-linking for protein interaction analysis of non-tagged integrin beta1.

ABSTRACT: Formaldehyde cross-linking of protein complexes combined with immunoprecipitation and mass spectrometry analysis is a promising technique for analysing protein-protein interactions, including those of transient nature. Here we used integrin beta1 as a model to describe the application of formaldehyde cross-linking in detail, particularly focusing on the optimal parameters for cross-linking, the detection of formaldehyde cross-linked complexes, the utility of antibodies, and the identification of binding partners. Integrin beta1 was found in a high molecular weight complex after formaldehyde cross-linking. Eight different anti-integrin beta1 antibodies were used for pull-down experiments and no loss in precipitation efficiency after cross-linking was observed. However, two of the antibodies

SUBMITTER: Klockenbusch C 

PROVIDER: S-EPMC2896913 | biostudies-literature | 2010

REPOSITORIES: biostudies-literature