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[资源] Elsevier 2014 新书：纯化及鉴定实验手册

书名 Experiments in the Purification and Characterization of Enzymes
--A Laboratory Manual
作者  Thomas Crowley / Jack Kyte
出版商  Elsevier -Academic Press
出版日期 Feb 27, 2014
页数  266
书号  ISBN: 978-0-12-409544-1
书评
Written in a logical, easy-to-understand manner,   Experiments in the Purification and Characterization of Enzymes: A Laboratory Manual is an indispensable resource for both students and instructors in the fields of biochemistry, molecular biology, chemistry, pharmaceutical chemistry, and related molecular life sciences such as cell biology, neurosciences, and genetics.
目录
Safety Guidelines for Biochemistry Laboratories
General Guidelines for Handling Solutions of Protein
Maintaining a Laboratory Notebook
Introduction to Enzymes Catalyzing Oxidation-Reductions with the Coenzyme NAD(P)
Computational Techniques for Biochemistry
Software for Analysis of Data
Computational Methods and Bioinformatics for Studying the Structure and Function of Proteins
Section 1:
FNR; Purification and Characterization of Ferredoxin-NADP+ Reductase from Chloroplasts of S. oleracea
1. Preparation of a Lysate of Chloroplasts and Sequestration of Proteins by an Anion-exchange Solid Phase;
2. Elution of Proteins from the Anion-exchange Solid Phase and Assays for the Concentration of Total Protein and FNR Activity;
3. Isolation of FNR by Adsorption and Elution
4. Electrophoresis of Proteins from the Chloroplasts and Purified FNR on Gels of Polyacrylamide Cast in Solutions of Dodecyl Sulfate;
5. Kinetic Assay for the Km of FNR for NADPH;
6. Estimation of the Size and Quaternary Structure of Native FNR Using Chromatography by Molecular Exclusion;
7. Detection of FNR in Extracts from Chloroplasts of Spinach by Immunoblotting
Section 2:
LuxG; Purification and Characterization of a Recombinant FMN Reductase from P. leiognathi
1. Expression of LuxG-hexahistidine in E. coli and Its Release from the Cells by Lysis;
2. Affinity Adsorption of LuxG-hexahistidine to a Solid Phase to which Ni2+ is Chelated;
3. Electrophoresis of the Polypeptides in the Lysate and Purified LuxG-hexahistidine;
4. Assay for the Flavin Reductase Activity of LuxG-hexahistidine;
5. Determination of the Michaelis Constant of LuxG-hexahistidine for NADH;
6. Determination of the Molar Concentration of LuxG-hexahistidine Spectrophotometrically
Section 3:
LDH; Purification and Characterization of Bovine LDH
1. Preparation of the Isoenzymes of LDH from Extracts of Various Tissues;
2. Spectrophotometric Assay of the Activity of Bovine LDH;
3. Separation and Visualization of the Isoenzymes of LDH by Electrophoresis on Cellulose Acetate;
4. Fractionation of Proteins in Bovine Heart by Precipitation with Sulfate;
5. Purification of LDH from Heart by Affinity Adsorption and Elution;
6. Concentration of Protein by the Bradford Stain and Absorbance at 280 nm;
7. Electrophoresis of Native Proteins from Bovine Heart and Purified LDH on a Gel of Polyacrylamide;
8. Electrophoresis of Unfolded Polypeptides from Bovine Heart and Purified LDH Saturated with Dodecyl Sulfate;
9. Determination of the Michaelis Constant, KmNADH, of the Isoenzymes of LDH for NADH;
10. Chromatography by Molecular Exclusion to Evaluate the Quaternary Structure of LDH;
11. Detection of LDH in Extracts of Bovine Tissue by Immunoblotting
Section 4:
Experimental Design;
1. Association of FNR with the Membranes of the Thylakoids in the Chloroplasts from Spinach;
2. Search for Multiple Flavin Reductases in P. leiognathi;
3. Assay for LDH Activity Specific to (R)-lactate in Bovine Mitochondria
Appendices
I. Measurement of Absorbance with Multiwell Plates and an Automated Plate Reader;
II. Operation of the Spectronic 20D+ Spectrophotometer
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2014-03-12 01:18:16, 8.09 M