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Enzyme unit definitions and assay design

 

 
 
 

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This guide explains key concepts such as ‘enzyme units’ and ‘enzyme activity’ in simple terms and discusses enzyme assay design and the importance of operating in the ‘linear range’. There are also some tips on setting up assay controls and subtracting blanks are discussed.

This guide covers:

  • Definitions of enzyme units
  • What is ‘enzyme activity’
  • Factors affecting enzyme activity
  • Developing assays and the importance of linear range
  • Assay time and temperature
  • Assay volume/sensitivity
  • Continuous assays (measuring product appearance over time)
  • What substrate concentration should I use?
  • Standard curves
  • Do I plot concentration or absolute amount on the x axis?
  • Controls and subtraction of ‘blank’ data
  • Calculating activity values

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