Argonaute (AGO) proteins play a key role in RNA silencing mechanisms. RNA silencing affects both RNA degradation and translation. The characterization of translation-associated RNA silencing mechanisms and components often requires polysome isolation and analysis. In this chapter, we describe the identification of AGO1 association with polysomes through polysome fractionation on sucrose gradient, preparation of proteins by filtration and concentration, and immunoblotting.
Keywords: Arabidopsis thaliana; Argonaute; Immunoblotting; MicroRNA; Polysomes; RNA silencing; Sucrose gradients.