Protein Analysis and Purification: Benchtop Techniques

Forsideomslag
Springer Science & Business Media, 22. dec. 2006 - 520 sider
How one goes about analyzing proteins is a constantly evolving ?eld that is no longer solely the domain of the protein biochemist. Inves- gators from diverse disciplines ?nd themselves with the unanticipated task of identifying and analyzing a protein and studying its physical properties and biochemical interactions. In most cases, the ultimate goal remains understanding the role(s) that the target protein is playing in cellular physiology. It was my intention that this manual would make the initial steps in the discovery process less time consuming and less intimidating. This book is not meant to be read from cover to cover. The expanded Table of Contents and the index should help locate what you are seeking. My aim was to provide practically oriented information that will assist the experimentalist in benchtop problem solving. The appendices are ?lled with diverse information gleaned from catalogs, handbooks, and manuals that are presented in a distilled fashion designed to save trips to the library and calls to technical service representatives. The user is encouraged to expand on the tables and charts to ?t individual experimental situations. This second edition pays homage to the computer explosion and the various genome projects that have revolutionized how benchtop scienti?c research is performed. Bioinformatics and In silico science are here to stay. However, the second edition still includes recipes for preparing buffers and methods for lysing cells.
 

Indhold

An Overview of This Manual
1
Protein Structure
8
Tracking the Target Protein
24
Labeling Proteins Present at the Plasma Membrane
30
would like to thank Paula Callaghan for giving me the opportunity
43
Electrophoretic Techniques
63
Safety Considerations
79
that is no longer solely the domain of the protein biochemist Investi
80
Protein Database Searches
244
Identifying and Analyzing Posttranslational
259
Lectins as Tools for Carbohydrate Analysis
276
B Phosphorylation
278
from PVDF Membranes
298
Isoprenylation
305
Metabolic Labeling with Precursors of the GPI Structure
313
Chromatography
324

Fluorescence TwoDimensional Difference Gel Electrophoresis
91
Recovery of Proteins from the Gel
105
Getting Started with Protein Purification
118
Manipulating Proteins in Solution
136
Precipitation Techniques
143
Membrane Proteins
153
Transfer and Detection of Proteins
177
Enzymatic Detection Methods
199
Removing Unwanted Background Signal from Xray Film
205
Identification of the Target Protein
211
Chemical Cleavage
219
Microsequencing from PVDF Membranes
225
Preparation of Proteins for MS Analysis
231
Protein Identification by MS
238
E Hydrophobic Interaction Chromatography HIC
356
F Affinity Chromatography
363
Recombinant Protein Techniques
385
Expression Identification
396
Affinity Tags
402
A Safety Considerations
433
Purification of Antibody Using Protein A Affinity Columns
439
Nucleic Acids
451
F Centrifugation
465
G Proteases and Proteolytic Enzyme Inhibitors
480
H Radioactivity
490
Rosenberg
503
Index
511
Copyright

Andre udgaver - Se alle

Almindelige termer og sætninger

Bibliografiske oplysninger