Measurement of inositol (1,4,5) trisphosphate in plant tissues by a competitive receptor binding assay

Abstract

The phosphoinositide signaling pathway is important for plant responses to many different stresses. As part of the responses to a stimulus, InsP 3 levels may increase rapidly and transiently. The receptor binding assay for InsP3 described here is easy to use and an ideal method to monitor and compare InsP3 levels in multiple samples from large scale experiments. The method is based on competitive binding of InsP3 to the mammalian brain InsP3 specific receptor protein. This chapter describes a protocol for extracting and neutralizing plant samples and performing the receptor binding assay (using a commercially available kit). The protocol described has been used effectively to monitor InsP3 levels in plant tissues of different origin and in response to different stresses. © 2013 Springer Science+Business Media, LLC.