Lipase Activity, Purification, and Expression -- Phospholipase A2 and Phosphatidylinositol-Specific Phospholipase C Assays by HPLC and TLC with Fluorescent Substrate -- Fluorometric Phospholipase Assays Based on Polymerized Liposome Substrates -- Triglyceride Lipase Assays Based on a Novel Fluorogenic Alkyldiacyl Glycerol Substrate -- Purification and Assay of Mammalian Group I and Group IIa Secretory Phospholipase A2 -- Determination of Plasmalogen-Selective Phospholipase A2 Activity by Radiochemical and Fluorometric Assay Procedures -- Human Plasma Platelet-Activating Factor Acetylhydrolase -- Assays for Pancreatic Triglyceride Lipase and Colipase -- Bile Salt-Activated Lipase -- Determining Lipoprotein Lipase and Hepatic Lipase Activity Using Radiolabeled Substrates -- Lysosomal Acid Lipase -- Hormone-Sensitive Lipase and Neutral Cholesteryl Ester Lipase -- Lecithin-Cholesterol Acyltransferase -- Purification of Lipases and Phospholipases by Heparin-Sepharose Chromatography -- Large-Scale Lipoprotein Lipase Purification from Adipose Tissue -- Purification of Rat Hepatic Lipase Essentially Free of Apolipoprotein E and Apolipoprotein B -- High-Level Expression and Purification of Human Hepatic Lipase from Mammalian Cells -- High-Level Baculoviral Expression of Hormone-Sensitive Lipase -- High-Level Baculoviral Expression of Lysosomal Acid Lipase -- One-Step Purification and Biochemical Characterization of Recombinant Pancreatic Lipases Expressed in Insect Cells -- Modulation of the Expression Level of Human Acidic Lipases by Various Signal Peptides -- Biochemical Characterization -- Immunodetection of Lipoprotein Lipase: Antibody Production, Immunoprecipitation, and Western Blotting Techniques -- Immunological Characterization of Digestive Lipases -- Determining Lipase Subunit Structure by Sucrose Gradient Centrifugation -- Detecting Ligands Interacting with Lipoprotein Lipase -- Monolayer Techniques for Studying Lipase Kinetics -- Efficient Immobilization of Phospholipase A2 -- Molecular Genetic Characterization -- An Introduction to Internet Resources for the Molecular and Genetic Analysis of the Lipases -- Techniques for the Measurement of Lipoprotein Lipase Messenger RNA -- In Vitro Transcription and Translation of Lipoprotein Lipase -- Induced Lipase Mutations in the Mouse.

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.