TY  - JOUR
TI  - Ex Vivo Measurement of Stable Isotope-Labeled Fatty Acid Incorporation Into Phospholipids in Isolated Mice Muscle
AU  - Sato, Tomoki 
AU  - Miura, Shinji 
VL  - 15
IS  - 7
PY  - 2025
DA  - 2025/04/05
SP  - e5257
C1  - Bio-protocol 2025;15:e5257
DO  - 10.21769/BioProtoc.5257
UR  - https://doi.org/10.21769/BioProtoc.5257
AB  - With the advancement of liquid chromatography–mass spectrometry (LC–MS/MS), the quantification of glycerophospholipid (PL) molecules has become more accessible, leading to the discovery of numerous enzymes responsible for determining the acyl groups attached to these molecules. Metabolic tracer experiments using radioisotopes and stable isotopes are powerful tools for defining the function of metabolic enzymes and metabolic flux. We have established an ex vivo muscle experimental system using stable isotope–labeled fatty acids to evaluate fatty acid incorporation into PL molecules. Here, we describe a method to incorporate fatty acids with stable isotope labels into excised skeletal muscle and detect the PL molecules containing labeled acyl chains by LC–MS/MS.
KW  - Stable isotope tracer
KW  - Phospholipid
KW  - Liquid chromatography–mass spectrometry
KW  - Skeletal muscle
KW  - Free-fatty acid
KW  - Acyl chain
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False