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One-step Purification of Phosphinothricin Acetyltransferase Using Reactive Dye-Affinity Chromatography

One-step Purification of Phosphinothricin Acetyltransferase Using Reactive Dye-Affinity Chromatography

Cunxi Wang, Thomas C Lee, Kathleen S Crowley, Erin Bell

Methods Mol Biol. 2015;1286:35-42.

PMID: 25749943

Abstract:

Reactive dye purification is an affinity purification technique offering unique selectivity and high purification potential. Historically, purification of phosphinothricin acetyltransferase (PAT) has involved several steps of precipitation and column chromatography. Here, we describe a novel purification method that is simple, time-saving, inexpensive, and reproducible. The novel method employs a single chromatography step using a reactive dye resin, Reactive brown 10-agarose. Reactive brown 10 preferentially binds the PAT protein, which can then be specifically released by one of its substrates, acetyl-CoA. Using Reactive brown 10-agarose, PAT protein can be purified to homogeneity from E. coli or plant tissue with high recovery efficiency.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42410994	Reactive Brown 10−Agarose			Price

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