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  • Mapping the lipolytic proteome of adipose tissue using fluorescent suicide inhibitors.

Mapping the lipolytic proteome of adipose tissue using fluorescent suicide inhibitors.

Methods in molecular biology (Clifton, N.J.) (2009-09-19)
Maximilian Schicher, Manfred Kollroser, Albin Hermetter
ABSTRACT

Lipases are responsible for the hydrolysis of acylglycerols and cholesteryl esters in animals, plants, and microorganisms. In this chapter we describe a tool for the concomitant analysis of lipases in complex proteomes. For this purpose, the target enzymes are selectively and covalently labelled with fluorescent suicide inhibitors. Stable lipid-protein complexes are formed that are separated by gel electrophoresis and identified by mass spectrometry.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Lipase from Aspergillus niger, powder (fine), ~200 U/g
Sigma-Aldrich
Lipase Substrate, for the titrimetric determination of enzyme activity
Sigma-Aldrich
Lipase from wheat germ, Type I, lyophilized powder, 5-15 units/mg solid
Sigma-Aldrich
Lipase from Candida rugosa, Type VII, ≥700 unit/mg solid
Sigma-Aldrich
Lipase from Rhizopus oryzae, powder (fine), ~10 U/mg
Sigma-Aldrich
Lipase Substrate, ≥95% (HPLC)