A malachite green-based assay to assess glucan phosphatase activity

Amanda R. Sherwood, Bradley C. Paasch, Carolyn A. Worby, Matthew S. Gentry

Research output: Contribution to journalArticlepeer-review

28 Scopus citations


With the recent discovery of a unique class of dual-specificity phosphatases that dephosphorylate glucans, we report an in vitro assay tailored for the detection of phosphatase activity against phosphorylated glucans. We demonstrate that, in contrast to a general phosphatase assay using a synthetic substrate, only phosphatases that possess glucan phosphatase activity liberate phosphate from the phosphorylated glucan amylopectin using the described assay. This assay is simple and cost-effective, providing reproducible results that clearly establish the presence or absence of glucan phosphatase activity. The assay described will be a useful tool in characterizing emerging members of the glucan phosphatase family.

Original languageEnglish
Pages (from-to)54-56
Number of pages3
JournalAnalytical Biochemistry
Issue number1
StatePublished - Apr 1 2013

Bibliographical note

Funding Information:
This work was supported by National Institutes of Health (NIH) Grants R00NS061803, P20RR020171, and R01NS070899 and by University of Kentucky College of Medicine start-up funds to M.S.G. This publication was also supported by Grant TL1 RR033172 from the National Center for Research Resources (NCRR), funded by the Office of the Director, NIH, and supported by the NIH Roadmap for Medical Research. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NCRR and NIH. We thank members from the Gentry lab and Dixon lab at the University of California, San Diego, including Ji Zhang, who supplied several enzymes used in this work in addition to providing helpful information in regard to their use.


  • Amylopectin
  • Glucan
  • Glycogen
  • Malachite green
  • Phosphatase
  • Starch

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology


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