Team:Washington/Protocols/Purified Enzyme Assay

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=Whole Cell Lysate Assay=
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<!---------------------------------------PAGE CONTENT GOES BELOW THIS---------------------------------------->
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*'''Assay'''
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**Add 90uL of 5uM substrate (in NaAc pH 4.0 buffer) to each well of a black fluorescent microtiter assay plate
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**Start reaction by adding 0.0125mg/mL enzyme (try to avoid bubbles and pippette quickly, but accurately)
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***Use the P20 multichannel with a taped cardboard stopper to make sure you don't hit the pellet!
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**Monitor the reaction with the SpectraMax
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<!---------------------------------------PAGE CONTENT GOES ABOVE THIS---------------------------------------->
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'''&larr; [[Team:Washington/Protocols|Back to Protocols]]'''
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&nbsp; &nbsp; &nbsp;
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</div>

Revision as of 01:11, 23 September 2011


Whole Cell Lysate Assay


  • Assay
    • Add 90uL of 5uM substrate (in NaAc pH 4.0 buffer) to each well of a black fluorescent microtiter assay plate
    • Start reaction by adding 0.0125mg/mL enzyme (try to avoid bubbles and pippette quickly, but accurately)
      • Use the P20 multichannel with a taped cardboard stopper to make sure you don't hit the pellet!
    • Monitor the reaction with the SpectraMax



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