Search our Test Catalog. Specify what to search (name, test code, titles, everywhere) and how to search (begins with, contains)

Unit Code 81927:
Fatty Acid Oxidation Probe Assay, Fibroblast Culture

Print Friendly View
Print Page Email Page

Method Description

Skin fibroblasts are incubated with cell medium enriched with palmitic

acid (C16:0 fatty acid), L-carnitine, and isotopically labeled L-valine

([13]C-Val) and L-isoleucine ([13]C-Ile). Cell lines deficient of 1 of the

enzymes involved in fatty acid oxidation and branched chain amino

acid metabolism fail to metabolize acyl-CoA species, which accumulate

in the cell medium as acylcarnitines. The medium is separated from

the cells following the incubation. The cell pellet is used for protein

determination and the medium will be spotted and dried on filter paper.

An acylcarnitine analysis is performed by tandem mass spectometry

(MS/MS) using a 1/4" filter paper punch, following the addition of

isotopically labeled acylcarnitines as internal standards, extraction and

derivatization to methyl esters. The assay is performed in triplicate.

(Matern D:  Acylcarnitines, incl. in vitro loading tests. In Laboratory Guide

to the Methods in Biochemical Genetics. Edited by N Blau, M Duran,

KM Gibson. Springer-Verlag, Heidelberg, 2006; in press)

Performing Laboratory Location

Rochester

Key