Principles and techniques of practical biochemistry fifth editionPDF|Epub|txt|kindle电子书版本网盘下载

Principles and techniques of practical biochemistry fifth editionPDF格式电子书版下载

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1 General principles of biochemical investigations&I.SIMPKINS1

1.1 The nature of biochemistry1

1.2 Bioenergetics4

1.3 Methods for investigating metabolism21

1.4 Practical considerations28

1.5 In vivomodels42

1.6 Invitromodels44

1.7 Microscopy66

1.8 Key terms75

1.9 Calculations76

1.10 Suggestions for further reading78

2 Molecular biology and basic techniques&R.RAPLEY80

2.1 Introduction80

2.2 Components and primary structure of nucleic acids80

2.3 Genes and genome complexity87

2.4 The nature of the genetic code90

2.5 Cellular location of nucleic acids90

2.6 The cellular functions of DNA93

2.7 The manipulation of nucleic acids：basic tools and techniques103

2.8 Isolation and separation of nucleic acids105

2.9 Restriction mapping of DNA fragments110

2.10 Nucleic acid blotting methods111

2.11 Gene probe derivation113

2.12 Labelling DNA gene probe molecules114

2.13 The polymerase chain reaction116

2.14 Nucleotide sequencing of DNA125

2.15 Bioinformatics and the Internet131

2.16 Key terms134

2.17 Calculations135

2.18 Suggestions for further reading136

3 Molecular cloning and gene analysis&R.RAPLEY138

3.1 Introduction138

3.2 Constructing gene libraries138

3.3 Cloning vectors148

3.4 Hybridisation and gene probes167

3.5 Screening gene libraries167

3.6 Applications of gene cloning171

3.7 Expression of foreign genes178

3.8 Analysing genes and gene expression184

3.9 Analysing whole genomes196

3.10 Molecular biotechnology and applications202

3.11 Key terms204

3.12 Suggestions for further reading205

4 Immunochemical techniques&R.THORPE AND S.THORPE206

4.1 Introduction206

4.2 Production of antibodies211

4.3 Purification and fragmentation of immunoglobulins222

4.4 Immunoprecipitation229

4.5 Labelling antibodies234

4.6 Immunoblotting240

4.7 Immunoassays244

4.8 Immunohisto/cytochemistry254

4.9 Affinity and avidity260

4.10 Immunochemical use of surface plasmon resonance260

4.11 Key terms261

4.12 Calculation262

4.13 Suggestions for further reading262

5 Centrifugation techniques&A.GRIFFITHS263

5.1 Introduction263

5.2 Basic principles of sedimentation264

5.3 Centrifuges and their uses271

5.4 Design and care of preparative rotors276

5.5 Sample containers284

5.6 Separation methods in preparative ultracentrifuges285

5.7 Performing density gradient separations290

5.8 Selection，efficiency and applications of preparative rotors296

5.9 Analysis of subcellular fractions300

5.10 Some applications of the analytical ultracentrifuge302

5.11 Safety aspects in the use of centrifuges305

5.12 Key terms306

5.13 Calculations306

5.14 Suggestions for further reading311

6 Protein structure，purification and characterisation&J.WALKER312

6.1 Ionic properties of amino acids and proteins312

6.2 Protein structure316

6.3 Protein purification318

6.4 Protein structure determination338

6.5 Key terms353

6.6 Calculations354

6.7 Suggestions for further reading356

7 Biomolecular interactions：Ⅰ Enzymes&K.WILSON357

7.1 Receptor-ligand binding357

7.2 Enzymes：characteristics and nomenclature358

7.3 Enzyme steady-state kinetics359

7.4 Enzyme assays378

7.5 Substrate assays385

7.6 Enzyme pre-steady-state kinetics386

7.7 Enzyme active sites and catalytic mechanisms389

7.8 Immobilised enzymes394

7.9 Cellular control of metabolic activity396

7.10 Key terms400

7.11 Calculations401

7.12 Suggestions for further reading402

8 Biomolecular interactions：Ⅱ Cell surface receptors and transporters&K.WILSON403

8.1 Cell surface receptor classification403

8.2 Quantitative aspects of receptor-ligand binding406

8.3 Receptor structures417

8.4 Mechanisms of signal transduction420

8.5 Signal amplification430

8.6 Key terms432

8.7 Membrane transport processes432

8.8 Physical diffusion433

8.9 Facilitated transport435

8.10 Active transport and ion channels438

8.11 Receptor-mediated endocytosis445

8.12 Key terms449

8.13 Calculations449

8.14 Suggestions for further reading451

9 Spectroscopic techniques：Ⅰ Atomic and molecular electronic spectroscopy&D.B.GORDON453

9.1 Introduction453

9.2 γ-Ray spectroscopy and γ-ray resonance spectroscopy456

9.3 X-ray spectroscopy458

9.4 Ultraviolet and visible light spectroscopy459

9.5 Spectrofluorimetry471

9.6 Circular dichroism spectroscopy479

9.7 Turbidimetry and nephelometry482

9.8 Luminometry483

9.9 Atomic spectroscopy485

9.10 Lasers490

9.11 Key terms491

9.12 Calculations492

9.13 Suggestions for further reading497

10 Spectroscopic techniques：Ⅱ Vibrational spectroscopy and electron and nuclear spin orientation in magnetic fieldsz&D.B.GORDON498

10.1 Introduction498

10.2 Infrared and Raman spectroscopy498

10.3 Electron spin resonance spectroscopy501

10.4 Nuclear magnetic resonance spectroscopy508

10.5 Key terms525

10.6 Suggestions for further reading526

11 Mass spectrometric techniques&D.B.GORDON527

11.1 Introduction527

11.2 The mass spectrometer527

11.3 Electron impact ionisation529

11.4 Chemical ionisation538

11.5 Field ionisation539

11.6 Ion desorption methods539

11.7 Ion evaporation methods551

11.8 Analysers555

11.9 Detectors564

11.10 Tandem mass spectrometry566

11.11 Key terms573

11.12 Calculations573

11.13 Suggestions for further reading579

12 Electrophoretic techniques&J.M.WALKER580

12.1 General principles580

12.2 Support media584

12.3 Electrophoresis of proteins588

12.4 Electrophoresis ofnucleic acids607

12.5 Capillary electrophoresis612

12.6 Key terms617

12.7 Calculation618

12.8 Suggestions for further reading618

13 Chromatographic techniques&K.WILSON619

13.1 Introduction619

13.2 Chromatography theory and practice623

13.3 Low pressure column chromatography631

13.4 High performance liquid chromatography637

13.5 Adsorption chromatography647

13.6 Partition chromatography649

13.7 Ion-exchange chromatography656

13.8 Molecular exclusion（permeation）chromatography661

13.9 Affinity chromatography665

13.10 Gas-liquid chromatography672

13.11 Thin-layer（planar）chromatography678

13.12 Selection of a chromatographic system681

13.13 Key terms682

13.14 Calculations683

13.15 Suggestions for further reading686

14 Radioisotope techniques&R.J.SLATER687

14.1 The nature of radioactivity687

14.2 Detection and measurement of radioactivity693

14.3 Other practical aspects of counting radioactivity and analysis of data713

14.4 Inherent advantages and restrictions of radiotracer experiments717

14.5 Safety aspects718

14.6 Applications of radioisotopes in the biological sciences721

14.7 Key terms726

14.8 Calculations726

14.9 Suggestions for further reading728

15 Electrochemical techniques&P.K.ROBINSON729

15.1 Introduction729

15.2 Principles of electrochemical techniques734

15.3 Redox reactions742

15.4 The pH electrode745

15.5 Ion-selective and gas-sensing electrodes748

15.6 The Clark oxygen electrode750

15.7 Electrochemical detectors for HPLC757

15.8 Biosensors760

15.9 Key terms768

15.10 Calculations768

15.11 Suggestions for further reading770

Index771