Bio-Rad Microplate Manager Software - Manual (Page 60)

Running Kinetic Assays
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Running Kinetic Assays
Running assays kinetically has many advantages:
• Kinetic assays are more accurate. The errors in endpoint assays such as the
time interval between substrate addition and reading in wells across the
plate, differences in the optical uniformity of wells, and the substrate
volume in the wells are eliminated.
• Using kinetics, assay sensitivity is greater for competitive assays.
• In kinetics the (acid) stop reagent and the decision to know when to stop the
enzyme reaction are eliminated.
• Kinetics is more accurate since rate, but not necessarily endpoint
absorbance, is directly proportional to the limiting reagent concentration.
• With kinetics, a broader concentration range is possible with fewer sample
dilutions.
• In kinetics, the assay time and substrate incubation are shorter.
Reading a Kinetic Assay
1. Click on the New Experiment icon or choose File > New Experiment.
2. Choose File > Instrument Setup.
3. Select Kinetic in the Reading Mode pop-up menu.
4. Select the desired kinetic parameter, Rate or Maximum Rate from the
Kinetic Mode popup.
In the kinetic mode, up to five different reading sets can be done successively. A
reading set consists of a number of times a plate is read, the delay between each read
and mix time, or before each reading.
To activate a reading set:
1. Click on the box next to the set number. Only sets that are checked will be
run.
2. Enter the number of reads in the Reads column.