Generation and Utilization of P450 Cholesterol Side‐Chain Cleavage Enzyme and 3β‐Hydroxysteroid Dehydrogenase Antibodies for Universal Detection

Abstract

The biosynthesis of steroids from steroidogenic cells are catalyzed by the two major enzymes, P450 side‐chain cleavage enzyme (P450scc) and 3β‐hydroxysteroid dehydrogenase (3β‐HSD). This article describes the construction of two novel polyclonal antibodies against conserved recombinant protein and the validation of these antibodies on fixed tissue sections of bovine corpus luteum. The polyclonal antibodies were used successfully in Western blots and specifically reacted with P450scc and 3β‐HSD protein in bovine luteal cell extracts. Thus, P450scc and 3β‐HSD are two specific polyclonal antibodies that are integral products in the investigation of the biological function and regulatory mechanism involved in steroidogenesis.

Keywords:

• Steroidogenesis
• P450scc
• 3β‐HSD
• Polyclonal antibody
• Bovine corpus luteum

ACKNOWLEDGMENT

The authors gratefully acknowledge the valuable technical advice of Dr. Guo, Department of Veterinary Medicine, National Taiwan University, on the plasmid construction.