Synthetic Protocol for AFCS: A Biologically Active Fluorescent Castasterone Analog Conjugated to an Alexa Fluor 647 Dye

Methods Mol Biol. 2017:1564:9-21. doi: 10.1007/978-1-4939-6813-8_2.

Abstract

Synthetic derivatization of hormonally active brassinosteroids (BRs) can provide useful small molecule tools to probe BR signaling pathways, such as fluorescent analogs. However, most biologically active BRs are not suitable for direct chemical conjugation techniques because their derivatization typically requires extensive synthetic work and chemistry expertise. Here, we describe an operationally simple, two-step procedure to prepare and purify an Alexa Fluor 647-castasterone (AFCS) from commercially available materials. The reported strategy is also amenable to the introduction of various other amine-based labeling groups.

Keywords: Brassinosteroids; Castasterone; Fluorescent labeling; Oxime conjugation; Steroid hormones.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Brassinosteroids / chemical synthesis*
  • Carbocyanines / chemistry
  • Chemistry Techniques, Synthetic*
  • Cholestanols / chemistry*
  • Chromatography, Thin Layer / methods
  • Fluorescent Dyes / chemistry
  • Oximes / chemistry*
  • Plant Growth Regulators / chemical synthesis*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Staining and Labeling / methods

Substances

  • Alexa Fluor 647
  • Brassinosteroids
  • Carbocyanines
  • Cholestanols
  • Fluorescent Dyes
  • Oximes
  • Plant Growth Regulators
  • castasterone