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Preparation of the membrane-permeant biarsenicals FlAsH-EDT2 and ReAsH-EDT2 for fluorescent labeling of tetracysteine-tagged proteins

Abstract

The membrane-permeant fluorogenic biarsenicals FlAsH-EDT2 and ReAsH-EDT2 can be prepared in good yields by a straightforward two-step procedure from the inexpensive precursor dyes fluorescein and resorufin, respectively. Handling of toxic reagents such as arsenic trichloride is minimized so the synthesis can be carried out in a typical chemistry laboratory, usually taking about 2–3 d. A wide range of other biarsenical reagents and intermediates that also bind to tetracysteine-tagged (CysCysProGlyCysCys) proteins can be prepared similarly using this general procedure.

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Figure 1: Synthesis of FlAsH-EDT2 and ReAsH-EDT2.
Figure 2: Structures of some other useful biarsenicals reagents and intermediates.
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Correspondence to Stephen R Adams or Roger Y Tsien.

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Adams, S., Tsien, R. Preparation of the membrane-permeant biarsenicals FlAsH-EDT2 and ReAsH-EDT2 for fluorescent labeling of tetracysteine-tagged proteins. Nat Protoc 3, 1527–1534 (2008). https://doi.org/10.1038/nprot.2008.144

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