Lipase and Phospholipase Protocols

A unique collection of step-by-step methods for the characterization, purification, and expression of mammalian lipases and phospholipases. The methods include the preparation of a variety of unique substrates for the determination of lipolytic activity, the isolation of purified lipase preparations, and the development of high expressing recombinant systems. There are also methods for the immunodectection and production of immunological reagents and the analysis of biochemical properties, such as subunit size, lipase kinetics, enzyme immobilization, and ligand interactions. The protocols covers more than a dozen specific enzymes, and provide well-established methods that can be adapted to the discovery of new lipase enzymes.Lipase and Phospholipase Protocols is an important new compilation of cutting-edge techniques and strategies for the characterization, purification, and expression of mammalian lipases and phospholipases. The methods include the preparation of a variety of radiolabeled and fluorescent substrates for the determination of lipolytic activity, proven strategies for the expression and purification of recombinant lipases and phospholipases, and the characterization of lipases by such techniques as immunodetection, subunit size determination, lipase kinetics, and ligand interactions. In addition, there are methods for the molecular and genetic analysis of the lipases, the quantification of mRNA levels, and in vitro-coupled transcription/translation. The protocols cover more than a dozen specific enzymes, and many can be applied directly to all lipases and phospholipases.§Lipase and Phospholipase Protocols constitutes a highly useful single-volume handbook of basic procedures and strategies to investigate this unique class of enzymes. Its step-by-step format works perfectly at the laboratory bench or by the computer, providing practical information and examples that will lead to successful design and execution of specific techniques. It not only furnishes a technical framework for the investigation of known lipases and phospholipases, but provides well-established methods readily daptable to the discovery of new lipase enzymes.