Accéder au contenu
MilliporeSigma

Simple and rapid real-time monitoring of LPL activity in vitro.

MethodsX (2020-04-11)
Stefan Kluge, Lisa Boermel, Martin Schubert, Stefan Lorkowski
RÉSUMÉ

Since elevated plasma triglycerides are an independent risk factor for cardiovascular diseases, lipoprotein lipase (LPL) is an interesting target for drug development. However, investigation of LPL remains challenging, as most of the commercially available assays are limited to the determination of LPL activity. Thus, we focused on the evaluation of a simple in vitro real-time fluorescence assay for the measurement of LPL activity that can be combined with additional cell or molecular biological assays in the same cell sample. Our procedure allows for a more comprehensive characterization of potential regulatory compounds targeting the LPL system. The presented assay procedure provides several advantages over currently available commercial in vitro LPL activity assays:1.12-well cell culture plate design for the simultaneous investigation of up to three different compounds of interest (including all assay controls).2.24 h real-time acquisition of LPL activity for the identification of the optimal time point for further measurements.3.Measurement of LPL activity can be supplemented by additional cell or molecular biological assays in the same cell sample.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
Milieu RPMI-1640, With L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
PMA, for use in molecular biology applications, ≥99% (HPLC)
Sigma-Aldrich
Milieu RPMI-1640, Modified, with sodium bicarbonate, without L-glutamine and phenol red, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Orlistat, ≥98%, solid
Sigma-Aldrich
L-Glutamine–Penicillin–Streptomycin solution, with 200 mM L-glutamine, 10,000 U penicillin and 10 mg steptomycin/mL in 0.9% NaCl, 0.1 μm filtered, BioReagent, suitable for cell culture