{"product_id":"fundamental-laboratory-approaches-for-biochemistry-and-biotechnology-isbn-9780470087664","title":"Fundamental Laboratory Approaches for Biochemistry and Biotechnology","description":"Ninfa\/Ballou\/Benore is a solid biochemistry lab manual, dedicated to developing research skills, allowing students to learn techniques and develop the the critical thinking and organizational approaches necessary to conduct laboratory research.   \u003cp\u003eNinfa\/Ballou\/Benore focuses on basic biochemistry laboratory techniques but also includes molecular biology exercises, a reflection of most courses which concentrate on traditional biochemistry experiments and techniques.  The experiments are designed so that theory and technique are learned as fundamental research tools, and the biochemistry and molecular biology applications are seamlessly integrated throughout the manual. The manual also includes an introduction to ethics in the laboratory, uncommon in similar manuals.  Most importantly, perhaps, is the authors’ three-pronged approach to encouraging students to think like a research scientist: first, the authors introduce the scientific method and the hypothesis as a framework for developing conclusive experiments; second, the manual’s experiments are designed to become increasingly complex in order to teach more advanced techniques and analysis; finally, gradually, the students are required to devise their own protocols.   In this way, students and instructors are able to break away from a “cookbook” approach and to think and investigate for themselves.\u003c\/p\u003e \u003cp\u003eSuitable for lower-level and upper-level courses; Ninfa spans these courses and can also be used for some first-year graduate work.\u003c\/p\u003e \u003cp\u003ePreface xv\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 1: Getting Started in Scientific Research 1\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e1.1 The Difference Between Experiments and Demonstrations 1\u003c\/p\u003e \u003cp\u003e1.2 Philosophy and Design of Experiments 3\u003c\/p\u003e \u003cp\u003e1.3 Designing Informative Experiments 4\u003c\/p\u003e \u003cp\u003e1.4 Ethics in Science 5\u003c\/p\u003e \u003cp\u003e1.5 Keeping a Laboratory Notebook 9\u003c\/p\u003e \u003cp\u003e1.6 Laboratory Reports 13\u003c\/p\u003e \u003cp\u003e1.7 Presentation and Analysis of Data 15\u003c\/p\u003e \u003cp\u003e1.8 The Minisymposium 18\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 2: Basic Procedures in the Biochemistry Laboratory 21\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e2.1 Laboratory Safety 21\u003c\/p\u003e \u003cp\u003e2.2 Special Safety Procedures are Required for Using Radioactive Materials or Operating the Autoclave 26\u003c\/p\u003e \u003cp\u003e2.3 Measurement of Weights, Volumes, and pH 30\u003c\/p\u003e \u003cp\u003e2.4 Various Instruments Used 38\u003c\/p\u003e \u003cp\u003e2.5 Other General Techniques 45\u003c\/p\u003e \u003cp\u003e2.6 Solutions and Dilutions 50\u003c\/p\u003e \u003cp\u003e2.7 Buffers and pH 52\u003c\/p\u003e \u003cp\u003e2.8 Appendix Calculating Titration Curves for Polyprotic Acids and Other Multiple Binding Site Receptors 62\u003c\/p\u003e \u003cp\u003e2.9 Equipment Used in This Course 64\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 3: Spectroscopic Methods 65\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e3.1 Introduction 65\u003c\/p\u003e \u003cp\u003e3.2 Design and Properties of Spectrophotometers 69\u003c\/p\u003e \u003cp\u003e3.3 Effects of Spectral Bandpass and Stray Light 72\u003c\/p\u003e \u003cp\u003e3.4 Recording Spectrophotometers 74\u003c\/p\u003e \u003cp\u003e3.5 Fluorescence Spectroscopy 75\u003c\/p\u003e \u003cp\u003e3.6 Chromogenic and Fluorogenic Reactions Used for Analysis 83\u003c\/p\u003e \u003cp\u003e3.7 Other Spectroscopic Techniques 84\u003c\/p\u003e \u003cp\u003e3.8 Mass Spectrometry (MS) 94\u003c\/p\u003e \u003cp\u003eExperiments 3-1 to 3-4 99\u003c\/p\u003e \u003cp\u003eReagents Needed for Chapter 3 103\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 4: Quantification of Protein Concentration 105\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e4.1 Purposes of Protein Quantification 105\u003c\/p\u003e \u003cp\u003e4.2 Factors to Consider in Choosing an Assay 107\u003c\/p\u003e \u003cp\u003e4.3 Non-Colorimetric Procedures for Quantification of Proteins 108\u003c\/p\u003e \u003cp\u003e4.4 Colorimetric Procedures for Quantification of Proteins 110\u003c\/p\u003e \u003cp\u003eExperiment 4-1 115\u003c\/p\u003e \u003cp\u003eReagents Needed for Chapter 4 119\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 5: Chromatography 121\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e5.1 Introduction 121\u003c\/p\u003e \u003cp\u003e5.2 Gel-Filtration (Size Exclusion or Gel-Permeation) Chromatography 125\u003c\/p\u003e \u003cp\u003e5.3 Affinity Chromatography 133\u003c\/p\u003e \u003cp\u003e5.4 Ion-Exchange Chromatography 143\u003c\/p\u003e \u003cp\u003e5.5 Hydrophobic Interaction Chromatography 149\u003c\/p\u003e \u003cp\u003eExperiments 5-1 and 5-2 152\u003c\/p\u003e \u003cp\u003eReagents Needed for Chapter 5 160\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 6: Gel Electrophoresis of Proteins 161\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e6.1 Process of Electrophoresis 161\u003c\/p\u003e \u003cp\u003e6.2 Polyacrylamide Gels 163\u003c\/p\u003e \u003cp\u003e6.3 SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE) of Proteins 165\u003c\/p\u003e \u003cp\u003e6.4 Detection of Proteins in SDS-Polyacrylamide Gels 171\u003c\/p\u003e \u003cp\u003e6.5 Applications of SDS-PAGE 173\u003c\/p\u003e \u003cp\u003eExperiments 6-1 and 6-2 183\u003c\/p\u003e \u003cp\u003eReagents Needed for Chapter 6 190\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 7: Overview of Protein Purification 191\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e7.1 Introduction 191\u003c\/p\u003e \u003cp\u003e7.2 Development of a Suitable Assay Procedure 193\u003c\/p\u003e \u003cp\u003e7.3 Time, Temperature, and Yield 195\u003c\/p\u003e \u003cp\u003e7.4 Selection of the Best Source Material 195\u003c\/p\u003e \u003cp\u003e7.5 Solubilization of the Protein 198\u003c\/p\u003e \u003cp\u003e7.6 Initial Steps of Purification 199\u003c\/p\u003e \u003cp\u003e7.7 Developing a Series of High-Resolution Chromatographic Steps 201\u003c\/p\u003e \u003cp\u003e7.8 Methods Used to Change Buffer and Concentrate Protein Samples 204\u003c\/p\u003e \u003cp\u003e7.9 A Logical Series of Steps 205\u003c\/p\u003e \u003cp\u003e7.10 Storage of the Purified Protein 205\u003c\/p\u003e \u003cp\u003e7.11 The Protein Purification Table 206\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 8: Subcellular Fractionation 209\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e8.1 Introduction 209\u003c\/p\u003e \u003cp\u003e8.2 Structural Organization of Prokaryotic and Eukaryotic Cells 210\u003c\/p\u003e \u003cp\u003e8.3 Overview of Fractionation Protocols 213\u003c\/p\u003e \u003cp\u003eExperiment 8-1 220\u003c\/p\u003e \u003cp\u003eReagents Needed for Chapter 8 227\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 9: Isolation and Characterization of an Enzyme—Alkaline Phosphatase From Escherichia Coli 229\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e9.1 Objectives 229\u003c\/p\u003e \u003cp\u003e9.2 Introduction and Basic Principles 229\u003c\/p\u003e \u003cp\u003e9.3 Purification of Alkaline Phosphatase 233\u003c\/p\u003e \u003cp\u003e9.4 Characterization of Purified Alkaline Phosphatase 242\u003c\/p\u003e \u003cp\u003eAppendix 9-1 Assay of Alkaline Phosphatase 243\u003c\/p\u003e \u003cp\u003eReagents and Equipment Needed for Chapter 9 247\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 10: Enzyme Kinetics 251\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e10.1 Why Use Steady-State Kinetics? 251\u003c\/p\u003e \u003cp\u003e10.2 Steady-State Kinetics Principles 252\u003c\/p\u003e \u003cp\u003e10.3 The Significance of \u003ci\u003eK\u003c\/i\u003em and \u003ci\u003eV\u003c\/i\u003emax 255\u003c\/p\u003e \u003cp\u003e10.4 Graphical Analysis 257\u003c\/p\u003e \u003cp\u003e10.5 Competitive, “Noncompetitive, ” and “Uncompetitive” Inhibitors 260\u003c\/p\u003e \u003cp\u003eExperiments 10-1 to 10-3 267\u003c\/p\u003e \u003cp\u003eReagents Needed for Chapter 10 273\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 11: Ligand Binding 275\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e11.1 Ligand Binding is the Key to Most Biological Processes 275\u003c\/p\u003e \u003cp\u003e11.2 Analysis of Ligand Binding at Equilibrium 277\u003c\/p\u003e \u003cp\u003e11.3 Digression on Regression 283\u003c\/p\u003e \u003cp\u003e11.4 Effects of the Concentrations of \u003ci\u003eL \u003c\/i\u003eand \u003ci\u003eR \u003c\/i\u003e283\u003c\/p\u003e \u003cp\u003e11.5 Effects of Two Sites and Cooperative Behavior 285\u003c\/p\u003e \u003cp\u003e11.6 Analysis of the Kinetics of Ligand Binding 292\u003c\/p\u003e \u003cp\u003e11.7 Methods Used to Study Receptor-Ligand Interactions 295\u003c\/p\u003e \u003cp\u003eExperiments 11-1 and 11-2 302\u003c\/p\u003e \u003cp\u003eReagents Needed for Chapter 11 308\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 12: Enzymatic Methods of Analysis 309\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e12.1 Enzymatic Analysis of Substrates 309\u003c\/p\u003e \u003cp\u003e12.2 Assays for Enzymatic Activity 310\u003c\/p\u003e \u003cp\u003e12.3 Practical Considerations 311\u003c\/p\u003e \u003cp\u003e12.4 Coupled Assays 312\u003c\/p\u003e \u003cp\u003e12.5 Experiments with Pyridine Nucleotide-Requiring Enzymes 314\u003c\/p\u003e \u003cp\u003eExperiments 12-1 to 12-4 317\u003c\/p\u003e \u003cp\u003eReagents Needed for Chapter 12 328\u003c\/p\u003e \u003cp\u003eAppendix 12-1: Sample Calculations 330\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 13: Recombinant DNA Techniques 337\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e13.1 Introduction 337\u003c\/p\u003e \u003cp\u003e13.2 Properties of Nucleic Acids 339\u003c\/p\u003e \u003cp\u003e13.3 Strategy of Recombinant DNA Techniques 341\u003c\/p\u003e \u003cp\u003e13.4 Cutting and Splicing DNA 343\u003c\/p\u003e \u003cp\u003e13.5 Gel Electrophoresis of DNA 347\u003c\/p\u003e \u003cp\u003e13.6 Introducing DNA into Cells 353\u003c\/p\u003e \u003cp\u003e13.7 Identifying Transformed Cells 354\u003c\/p\u003e \u003cp\u003e13.8 Vectors, Hosts, and Libraries for Recombinant DNA Experiments 359\u003c\/p\u003e \u003cp\u003e13.9 Applications of Recombinant DNA Technology 366\u003c\/p\u003e \u003cp\u003eExperiments 13-1 to 13-4 375\u003c\/p\u003e \u003cp\u003eReagents Needed for Chapter 13 386\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 14: Polymerase Chain Reaction (PCR) Technology 389\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e14.1 Introduction 389\u003c\/p\u003e \u003cp\u003e14.2 Principle of the PCR Method 390\u003c\/p\u003e \u003cp\u003e14.3 The Three Steps of PCR are Controlled by Temperature 392\u003c\/p\u003e \u003cp\u003e14.4 Applications of PCR 394\u003c\/p\u003e \u003cp\u003e14.5 Legal and Ethical Issues 410\u003c\/p\u003e \u003cp\u003eExperiments 14-1 to 14-6 412\u003c\/p\u003e \u003cp\u003eReagents Needed for Chapter 14 423\u003c\/p\u003e \u003cp\u003e\u003cb\u003eChapter 15: Using the Computer and the Internet for Biochemical Research and Communication 425\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e15.1 Introduction to the World Wide Web (the Internet) 425\u003c\/p\u003e \u003cp\u003e15.2 Information on the Internet Useful to Biochemists 431\u003c\/p\u003e \u003cp\u003e15.3 Literature and Data Searches 432\u003c\/p\u003e \u003cp\u003e15.4 Visualization of Molecular Structures 440\u003c\/p\u003e \u003cp\u003e15.5 Other Useful Information 442\u003c\/p\u003e \u003cp\u003e15.6 Homework Assignment 445\u003c\/p\u003e \u003cp\u003eIndex 447\u003c\/p\u003e \u003cb\u003eAlexander J. Ninfa\u003c\/b\u003e is Associate Professor in the Department of Biological Chemistry at the University of Michigan and is internationally recognized for his work on the biochemical mechanisms of signal transduction and transcriptional regulation in bacteria. \u003cp\u003e\u003cb\u003eDavid P. Ballou\u003c\/b\u003e is Professor in the Department of Biological Chemistry at the University of Michigan and is widely known for his work in rapid kinetics and the study of enzyme mechanisms involving redox coenzymes.\u003c\/p\u003e","brand":"Wiley","offers":[{"title":"Default Title","offer_id":47989254979813,"sku":"NP9780470087664","price":98.5,"currency_code":"USD","in_stock":false}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/1842\/7735\/files\/9780470087664.jpg?v=1761783397","url":"https:\/\/k12savings.com\/products\/fundamental-laboratory-approaches-for-biochemistry-and-biotechnology-isbn-9780470087664","provider":"K12savings","version":"1.0","type":"link"}