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

Alexis M Ceasrine; Lauren A Green; Staci D Bilbo

doi:10.1016/j.xpro.2022.101669

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

STAR Protoc. 2022 Dec 16;3(4):101669. doi: 10.1016/j.xpro.2022.101669. Epub 2022 Sep 13.

Authors

Alexis M Ceasrine¹, Lauren A Green², Staci D Bilbo³

Affiliations

¹ Department of Psychology and Neuroscience, Duke University, Durham, NC 27710, USA. Electronic address: alexis.ceasrine@duke.edu.
² Department of Psychology and Neuroscience, Duke University, Durham, NC 27710, USA.
³ Department of Psychology and Neuroscience, Duke University, Durham, NC 27710, USA; Departments of Neurobiology and Cell Biology, Duke University, Durham, NC 27710, USA. Electronic address: staci.bilbo@duke.edu.

Abstract

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 tissues. 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).

Keywords: Immunology; Neuroscience.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Biological Assay
Endotoxins* / analysis
Horseshoe Crabs*
Kinetics
Limulus Test / methods
Mice

Substances

Endotoxins

Abstract

Publication types

MeSH terms

Substances

Grants and funding