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Caspase-3/7 cell line screen for test compound

Caspase 3/7 activity is a cell based assay that measures apoptosis.  Commercially available reagents enable high-throughput screening of apoptosis inducing test compounds or contract research organizations can readily perform the compound screening.  A brief overview of the protocol is as follows:

  • Cells in logarithmic growth are seeded in a 96-well plate
  • Depending on the scope of the experiment, the test compound is added either as a single concentration or in a dilution series
  • Cells are harvested at various time-points after addition of the test compound (i.e. 6, 12, 24 and 48 hours)
  • In the appropriate well for each time-point, the caspase reagent is added to the cells, resulting in cell lysis and caspase cleavage of the fluorescent substrate
  • The resultant fluorescent signal is measured using a plate reader (i.e. Molecular Device Spectramax Gemini XPS plate reader)
  • The fluorescent signal is proportional to the amount of caspase activity present as a result of treatment by the test article

Altogen Labs, a contract research organization, perform the Caspase 3/7 apoptosis cell-based assay.  See here for more information.

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