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Method and reagent for monitoring apoptosis and distinguishing apoptosis from necrosis

  • US 5,976,822 A
  • Filed: 08/20/1997
  • Issued: 11/02/1999
  • Est. Priority Date: 05/18/1995
  • Status: Expired due to Fees
First Claim
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1. An assay reagent for determining the activity of an enzyme in a metabolically active whole cell, said assay reagent comprising at least one water soluble physiologically acceptable salt having the ability to pass through a cell membrane, said assay compound having an unblocked leaving group selected for cleavage by an enzyme to be analyzed selected from cysteine protease, dipeptyl peptidase and calpain, and a fluorogenic indicator group being selected for its ability to have a non-fluorescent first state when joined to the leaving group, and a fluorescent second state excitable at a wavelength above 450 nm when the unblocked leaving group is cleaved from the indicator group by the enzyme, wherein said fluorogenic indicator group is selected from the group consisting of rhodamine 110, rhodol, fluorescein and derivative thereof, said assay reagent having a fluorescence less than the auto-fluorescence of a metabolically active cell.

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