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Methods:Malachite green phosphatase activity

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To determine whether proteins have phosphatase activity. The assay detects the release of orthophosphate.

Materials Needed



User notes

Baykov et al. (1988) [1] describes the assay used by Kuznetsova et al. 2005 and 2006. [2][3]

In looking for this reference, I came across another methods paper.[4]


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  1. Baykov, AA et al. (1988) A malachite green procedure for orthophosphate determination and its use in alkaline phosphatase-based enzyme immunoassay. Anal. Biochem. 171 266-70 PubMed EcoliWiki page
  2. Kuznetsova, E et al. (2006) Genome-wide analysis of substrate specificities of the Escherichia coli haloacid dehalogenase-like phosphatase family. J. Biol. Chem. 281 36149-61 PubMed EcoliWiki page
  3. Kuznetsova, E et al. (2005) Enzyme genomics: Application of general enzymatic screens to discover new enzymes. FEMS Microbiol. Rev. 29 263-79 PubMed EcoliWiki page
  4. Geladopoulos, TP et al. (1991) A malachite green colorimetric assay for protein phosphatase activity. Anal. Biochem. 192 112-6 PubMed EcoliWiki page