Work

Protocol for the detection of proteolytic activity in female Aedes aegypti mosquito midgut extracts using fluorogenic AMC protease substrates

Year: 2025

Type: article

Abstract: Female Ae. aegypti mosquitoes utilize midgut proteolytic enzymes to digest blood meal proteins, but activity detection has largely depended on the use of chromogenic substrates. Here, we present a protocol for the detection of proteolytic activity in female mosquito midgut extracts using fluorescent 7-amino-4-methylcoumarin (AMC) protease substrates. We describe steps for midgut tissue and substrate preparation, as well as proper dilutions for easy fluorescent detectable readouts in 30 μL reactions. This approach saves time and minimizes sample usage. For complete details on the use and execution of this protocol, please refer to Ramirez et al. 1

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.103899

Authors: Abigail G. Ramirez, Daniel Fong, My Anh Le, Jun Isoe, Alberto A. Rascón

Institutions: Arizona State University, University of Arizona

Language: English

Topic: Mosquito-borne diseases and control

Subfield: Public Health, Environmental and Occupational Health

Field: Medicine

Domain: Health Sciences

Open Access status: gold

Estimated APC paid: $1,550

Funder: National Institute of General Medical Sciences