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A Colorimetric Microplate Assay Method for High Throughput Analysis of Lipase Activity

  • Choi, Suk-Jung (Department of Chemistry, Kangnung National University) ;
  • Hwang, Jung-Min (Department of Chemistry, Kangnung National University) ;
  • Kim, Sung-Il (Department of Chemistry, Kangnung National University)
  • Received : 2003.01.03
  • Accepted : 2003.02.04
  • Published : 2003.07.31

Abstract

The present work describes a colorimetric microplate assay for lipase activity based on the reaction between 5,5'-dithiobis(2-nitro benzoic acid) (DTNB) and the hydrolysis product of 2,3-dimercapto-1-propanol tributyrate (DMPTB). Reaction mixtures containing DTNB, DMPTB, and lipase were prepared in microplate wells, and the absorbance at 405nm was recorded after incubation at $37^{\circ}C$ for 30 min. A linear relationship was obtained in the range of 0.1-1 U of lipase activity by this method. The reaction conditions were also optimized for the range of 0.01-0.1 U or 1-10 U. When assaying crude tissue extracts, the reaction of DTNB with non-specific reducing agents created a major source of error. However, this error was corrected by the use of blank samples that did not contain DMPTB.

Keywords

Colorimetric;Lipase assay;Microplate

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