McClean:Western Blot

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Revision as of 08:28, 30 May 2013 by Ping Xu (talk | contribs) (Stock Solutions)
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This is a protocol to perform rapid yeast protein prep for SDS PAGE and Western.


  • Beta- mercaptoethanol
  • 4X LDS sample buffer(Invitrogen NP0008)
  • Protease inhibitor(Roche catlog number: 11836170001)
  • Phosphatase inhibitor(Fisher catlog number: 78420)
  • 20X NuPAGE running buffer(Invitrogen NP0001)
  • Prestained Protein Ladder(Invitrogen catlog number:10748-010)
  • Magic Marker(Invitrogen catlog number:LC5602)
  • NuPage pre-cast gel
  • PVDF membrane(Invitrogen catlog number:LC2005)
  • Methanol
  • 20X NuPAGE buffer(Invitrogen NP0006)
  • 1M Tris, pH 8.0
  • 2.5M NaCl
  • Tween 20
  • Nonfat dry milk
  • Primary and secondary antibody

Stock Solutions

Stock Solution 1

  • 4X LDS sample buffer(Invitrogen NP0008)

Stock Solution 2

  • 20X NuPAGE running buffer(Invitrogen NP0001)

Stock Solution 3

  • 1M Tris, pH 8.0

Stock Solution 4

  • * 2.5M NaCl


  1. Step 1
  2. Step 2
  3. Step 3


Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

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Gietz, R.D. and R.A. Woods. (2002) TRANSFORMATION OF YEAST BY THE Liac/SS CARRIER DNA/PEG METHOD. Methods in Enzymology 350: 87-96.


or instead, discuss this protocol.