Direct measurement of free and esterified cholesterol mass in differentiated human podocytes: A TLC and enzymatic assay-based method

Christopher E. Pedigo, Sandra M. Merscher, Alessia Fornoni

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Esterified cholesterol content is often lower than free cholesterol content in biological systems and thus the determination of the esterified cholesterol content of cells is often challenging. Traditional methods use enzymatic assays in which an indirect measurement of the esterified cholesterol content is obtained by subtracting the measurements of the free from the total cholesterol content. However, this approach fails in the case where the total cholesterol content of cells is unchanged while the ratio of free to esterified cholesterol changes such that total and free cholesterol content are very similar and thus the difference may fall within the background noise of the enzymatic assay. To overcome this challenge, we here describe a method that utilizes a TLC-based technique to isolate esterified cholesterol. Isolated esterified cholesterol can then be measured using traditional enzymatic methods. Therefore, this method provides a practical and more sensitive assay to measure esterified cholesterol content in cellular extracts.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages51-56
Number of pages6
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1609
ISSN (Print)1064-3745

Keywords

  • Cellular cholesterol
  • Esterified cholesterol content
  • Esterified cholesterol mass
  • Podocyte cholesterol

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'Direct measurement of free and esterified cholesterol mass in differentiated human podocytes: A TLC and enzymatic assay-based method'. Together they form a unique fingerprint.

Cite this