User:Christine Doan/Notebook/Identifying Kinase Substrates of PfCK2/2014/03/20

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new CK2

  • Sonicate 2x at 30% amplitude, 20sec on, 20sec off
  • Normalize concentrations
    • UI: 1ml culture in 500ul MK buffer, resuspend, take 10ul + 2.5ul 5x GLB
    • total I: 500ul culture in 500ul MK buffer, resuspend, take 20ul + 5ul 5x GLB
    • Pel: 5ml culture in 8ml MK buffer, finally resuspended, take 32ul + 8ul 5x GLB
    • Sup: 8ml total, take 32ul + 8ul 5x GLB
  • Run two sets on one gel: marker, UI, I, Pel, Sup
    • Stain one, semi-dry Western transfer the other
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