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Merck

A malachite green-based assay to assess glucan phosphatase activity.

Analytical biochemistry (2012-12-04)
Amanda R Sherwood, Bradley C Paasch, Carolyn A Worby, Matthew S Gentry
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Schaeffer and Fulton Spore Stain Solution A, for microscopy
Sigma-Aldrich
Malachite Green oxalate salt, Technical grade
Supelco
Malachite Green oxalate salt, VETRANAL®, analytical standard
Sigma-Aldrich
Malachite Green oxalate salt, for microscopy, crystalline, S. No.: 754
Sigma-Aldrich
Amylopectin from potato starch
Sigma-Aldrich
Malachite Green oxalate salt, certified by the Biological Stain Commission
Sigma-Aldrich
Amylopectin from maize
Sigma-Aldrich
Malachite Green solution, for microscopy