User:Tk/Notebook/MF/2008/09/12

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Clontech In-Fusion protein identification

  • Recycled In-Fusion 2.0 CF dry down kit from Drew's lab
  • Dissolve tube material in 50 ul water
  • Run 20 ul + 5 ul LDS loading buffer, heated to 70C
  • Run also 5x dilutions
  • No DTT or other reducing agent.
  • Bands at 120 and 63 kD
  • Also a very bright band at the well edge, probably unreduced protein
  • Run was with MES buffer, retry with MOPS buffer to give better separation.
  • Pre-run 30 min, run 1:30 at 150 V, about 13 watts