Unit

UNIT 10.15 Purification of Recombinant Proteins and Study of Protein Interaction by Epitope Tagging

  1. Ning Zhang,
  2. Jin-Long Chen

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1015s41

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Zhang, N. and Chen, J.-L. 2001. Purification of Recombinant Proteins and Study of Protein Interaction by Epitope Tagging. Current Protocols in Molecular Biology. 41:VI:10.15:10.15.1–10.15.9.

Author Information

  1. Tularik, Inc., South San Francisco, California

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JAN 1998

Abstract

A protein molecule can be engineered to include a short stretch of residues corresponding to an epitope to facilitate its subsequent biochemical and immunological analysis; a technique often referred to as “epitope tagging.” This unit presents a protocol for small-scale immunoprecipitation of epitope-tagged recombinant proteins expressed in transiently transfected mammalian cells. The immunoprecipitant can then be analyzed by SDS-PAGE. An immunoprecipitation protocol is also provided that has been optimized for use with a baculovirus overexpression system. An Alternate Protocol describes how multisubunit complexes can be assembled by starting with a core protein affixed to beads via an epitope tag, and adding the other members of the complex in a stepwise manner.