Protocol to measure endotoxin from opaque tissues in mice using an optimized kinetic limulus amebocyte lysate assay

Endotoxin accumulation has been widely noted in several pathologies ranging from metabolic dysregulation to bacterial infection. Using limulus amebocyte lysate (LAL) assays to detect endotoxin load has been the only reliable way to assess endotoxin accumulation, but assays optimized for detection in opaque tissues are still lacking. We optimized a sensitive Kinetic LAL assay for endotoxin detection from murine tis-sues. In this protocol, we describe tissue collection and homogenization, followed by the procedure to run the assay and data analysis.For complete details on the use and execution of this protocol, please refer to Ceasrine et al. (2022).

Automated summary

Endotoxin accumulation has been widely observed in various diseases, and the use of limulus amebocyte lysate (LAL) assays for endotoxin detection has been the most reliable method. However, there is still a lack of optimized assays for detecting endotoxin in opaque tissues. This study optimized a sensitive Kinetic LAL assay for endotoxin detection in murine tissues.