A Chromogenic Quantification of Protein Expression in Zebrafish Larvae

Abstract

Western blot is a versatile and widely used technique in many areas of molecular biology and biotechnology for studying different protein characteristics. In general, the Western blot technique involves the extraction of proteins from the samples such as cells or tissues, which, after denaturation, are separated by molecular size using electrophoresis. The protein is then transferred to a membrane, typically PVDF or nitrocellulose, which, after blocking, is probed with specific antibodies labeled with a detection agent. Overall, this allows the recognition and binding to the target protein allowing the visualization of bands, a step called immunodetection. Over the years, new approaches to the Western blotting technique have been proposed to overcome performance limitations.This chapter describes a routine procedure for protein evaluation in zebrafish (Danio rerio) larvae, a widely used animal model for predicting the toxicity of drugs, by using a chromogenic substrate and allowing the proper execution of the technique without the costly equipment needed for detection.

Keywords: Larvae; Protein; SDS-PAGE; TMB; Western blot; Zebrafish.