Immunoisolation of protein complexes from Xenopus

Frank L. Conlon, Yana Miteva, Erin Kaltenbrun, Lauren Waldron, Todd M. Greco, Ileana M. Cristea

Research output: Chapter in Book/Report/Conference proceedingChapter

13 Scopus citations

Abstract

The immunoaffinity isolation of protein complexes is an essential technique for the purification and -concentration of protein complexes from cells and tissues. In this chapter we present the methodologies for the purification of proteins and protein complexes from Xenopus laevis and Xenopus tropicalis. Specific to this protocol are the techniques for the cryolysis of Xenopus cells and tissues, a procedure that limits contamination from yolk proteins while preserving endogenous protein complexes, the methodologies for immunoaffinity purification of proteins using magnetic beads, and the protocols for western blot analysis. In addition, the procedures in this chapter can be extended to use with proteomic analysis of protein complexes as presented in the following chapter.

Original languageEnglish (US)
Title of host publicationXenopus Protocols
Subtitle of host publicationPost-Genomic Approaches
EditorsStefan Hoppler, Peter D. Vize
Pages369-390
Number of pages22
DOIs
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
Volume917
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Keywords

  • Cryolysis
  • Immunoaffinity purification
  • Immunoisolation
  • Immunoprecipitation
  • Protein complex
  • Protein extraction
  • Protein interactions
  • Tissue lysis
  • Western blot analysis
  • Xenopus

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  • Cite this

    Conlon, F. L., Miteva, Y., Kaltenbrun, E., Waldron, L., Greco, T. M., & Cristea, I. M. (2012). Immunoisolation of protein complexes from Xenopus. In S. Hoppler, & P. D. Vize (Eds.), Xenopus Protocols: Post-Genomic Approaches (pp. 369-390). (Methods in Molecular Biology; Vol. 917). https://doi.org/10.1007/978-1-61779-992-1-21