Immunoisolation of protein complexes from Xenopus

Methods Mol Biol. 2012;917:369-90. doi: 10.1007/978-1-61779-992-1_21.

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.

MeSH terms

  • Animals
  • Antibodies, Immobilized / chemistry
  • Antibodies, Immobilized / immunology
  • Antibody Affinity
  • Blotting, Western
  • Buffers
  • Electrophoresis, Polyacrylamide Gel
  • Embryo, Nonmammalian / metabolism
  • Immunoprecipitation / methods*
  • Magnets / chemistry
  • Microspheres
  • Multiprotein Complexes / immunology
  • Multiprotein Complexes / isolation & purification*
  • Xenopus Proteins / immunology
  • Xenopus Proteins / isolation & purification*
  • Xenopus laevis / metabolism*

Substances

  • Antibodies, Immobilized
  • Buffers
  • Multiprotein Complexes
  • Xenopus Proteins