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Merck

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

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

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.

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製品内容

Sigma-Aldrich
RPMI-1640培地, With L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
ホルボール 12-ミリスタート 13-アセタート, for use in molecular biology applications, ≥99% (HPLC), Molecular Biology
Sigma-Aldrich
RPMI-1640培地, Modified, with sodium bicarbonate, without L-glutamine and phenol red, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
オルリスタット, ≥98%, solid
Sigma-Aldrich
L-グルタミン–ペニシリン–ストレプトマイシン 溶液, 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