Improved elution conditions for native co-immunoprecipitation.

ABSTRACT:

Background

Native immunoprecipitation followed by protein A-mediated recovery of the immuno-complex is a powerful tool to study protein-protein interactions. A limitation of this technique is the concomitant recovery of large amounts of immunoglobulin, which interferes with down-stream applications such as mass spectrometric analysis and Western blotting. Here we report a detergent-based "soft" elution protocol that allows effective recovery of immunoprecipitated antigen and binding partners, yet avoids elution of the bulk of the immunoglobulin.

Methodology/principal findings

We assessed the performance of the soft elution protocol using immunoprecipitation of Adaptor protein complex 1 (AP-1) and associated proteins as a test case. Relative to conventional elution conditions

SUBMITTER: Antrobus R 

PROVIDER: S-EPMC3063181 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature