An improved 96-well turbidity assay for T4 lysozyme activity
XAVIER UNIV OF LOUISIANA NEW ORLEANS NEW ORLEANS United States
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T4 lysozyme T4L is an important model system for investigating the relationship between protein structure and function. Despite being extensively studied, a reliable, quantitative activity assay for T4L has not been developed. Here, we present an improved T4L turbidity assay as well as an affinity-based T4L expression and purification protocol. This assay is designed for 96-well format and utilizes conditions amenable for both T4L and other lysozymes. This protocol enables easy, efficient, and quantitative characterization of T4L variants and allows comparison between different lysozymes. Our method Is applicable for all lysozymes, with enhanced sensitivity for T4 lysozyme compared to other 96-well plate turbidity assays Utilizes standardized conditions for comparing T4 lysozyme variants and other lysozymes and Incorporates a simplified expression and purification protocol for T4 lysozyme.
- Test Facilities, Equipment and Methods