Overview

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.

Full Product Details

Author:   Mark Doolittle ,  Karen Reue
Publisher:   Humana Press Inc.
Imprint:   Humana Press Inc.
Edition:   1999 ed.
Volume:   109
Dimensions:   Width: 15.50cm , Height: 2.50cm , Length: 23.50cm
Weight:   0.739kg
ISBN:  

9780896035461

ISBN 10:   0896035468
Pages:   362
Publication Date:   01 October 1998
Audience:   College/higher education ,  Professional and scholarly ,  Undergraduate ,  Professional & Vocational
Format:   Hardback
Publisher's Status:   Active
Availability:   In Print  
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Table of Contents

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.

Reviews

...The book can be recommended to those scientists who are deeply engaged in lipid chemistry and analysis as well as enzymology. It provides a detailed listing of literature and other sources. Thus it can be successfully used as an introduction to new research fields and experimental techniques... -Nahrung In view of the rapid increase in the number of new lipases and phospholipases that have been recently identified, the focus of this book is timely...In this volume, modern, up-to-date protocols with sufficient detail for experienced or peripheral investigators in this area are provided, and should prove to be a valuable resource. -The Quarterly Review of Biology

...The book can be recommended to those scientists who are deeply engaged in lipid chemistry and analysis as well as enzymology. It provides a detailed listing of literature and other sources. Thus it can be successfully used as an introduction to new research fields and experimental techniques... - Nahrung In view of the rapid increase in the number of new lipases and phospholipases that have been recently identified, the focus of this book is timely In this volume, modern, up-to-date protocols with sufficient detail for experienced or peripheral investigators in this area are provided, and should prove to be a valuable resource. - The Quarterly Review of Biology

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