A rapid Western blotting protocol for the Xenopus oocyte

Cold Spring Harb Protoc. 2013 Mar 1;2013(3):10.1101/pdb.prot072793 pdb.prot072793. doi: 10.1101/pdb.prot072793.

Abstract

Often experimentalists require a quantitative assessment of the levels of heterologously expressed proteins to best interpret changed Ca(2+) signaling patterns. Here, we detail a rapid and convenient western blotting method for individual Xenopus oocytes. The method exploits recently introduced rapid blotting systems, commercially available from Invitrogen (iBlot) or Bio-Rad (Trans-Blot Turbo). The key advantage is speed: from live cell to transferred membrane in <1 h. Therefore, oocytes can be conveniently processed for western blotting to assess relative expression levels, even after a long day of Ca(2+) imaging experiments.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Blotting, Western / methods*
  • Molecular Biology / methods*
  • Oocytes / chemistry*
  • Recombinant Proteins / analysis*
  • Xenopus

Substances

  • Recombinant Proteins