SH2 Domains pp 163-172 | Cite as

Expression and Purification of Soluble STAT5b/STAT3 Proteins for SH2 Domain Binding Assay

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1555)

Abstract

When a large hydrophobic full-length protein is expressed in bacteria, it is often challenging to obtain recombinant proteins in the soluble fraction. One way to overcome this challenge is expression of deletion mutants that have improved solubility while maintaining biological activity. In this chapter, we describe a protocol for expression of truncated forms of STAT5b and STAT3 proteins that are soluble and retain SH2-mediated activity for phospho-Tyr peptide recognition.

Key words

STAT5b STAT3 SH2 domain Purification Soluble protein 

Notes

Acknowledgment

This work was supported by National Institute of Biomedical Innovation (NIBO) grant number 06–02, JSPS KAKENHI Grant Number 23510281/26430166, and Pharma Valley Center (PVC) Shizuoka, Japan.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Center for Drug Discovery, Graduate School of Pharmaceutical SciencesUniversity of ShizuokaSuruga-ku, ShizuokaJapan
  2. 2.Innovative Research DivisionMitsubishi Tanabe Pharma CorporationSaitamaJapan

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