User:Tk/Notebook/MF/2008/09/14

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Revision as of 15:45, 14 September 2008 by Tk (talk | contribs) (→‎In-Fusion protein identification)
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In-Fusion protein identification

  • Re-run In-Fusion gel
    • 10% Bis-tris gel
    • 150 V, 1 hr 20 min
    • prerun 30 min
    • dissolve In-Fusion mix in 13 ul water
    • Add to serial dilution mix of:
      • 2 ul beta-mercaptoethanol
      • 5 ul LDS sample buffer
    • 3 additional mixes of BME + LDS + 13 ul water
    • 2x serial dilutions into the LDS mix
    • incubate 15 min at 70C
    • run 10 ul SeeBlue plus 2 marker lanes
    • run 20 ul of each dilution
    • wash 2x with microwaved DI water 10 min
    • Add Simply Blue stain covering gel, incubate overnight
    • Wash 2x with hot DI water + kimwipes
    • Image with gel imager in water
    • store in plastic bag


128 pixel


  • There appears to be two proteins present
    • One is about 68 kD
    • A second, about 8x less in mass, appears to be about 155 kD




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