Protein Analysis and Purification: Benchtop TechniquesSpringer 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
1 | |
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 |
511 | |
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Almindelige termer og sætninger
activity adding addition affinity allow amino acid amount analysis antibody antigen band beads binding Biochem Biol blot bonds buffer cells centrifugation charge chromatography cleavage column Comments complex concentration containing database described detection detergent determine digestion dilute electrophoresis elution enzyme et al exchanger expression extract Figure final flow fraction function fusion protein gene glycosylation gradient groups hydrophobic identified increase incubate interactions interest isolated labeled mass Materials membrane method mixture modifications molecular weight molecules peptide performed phase phosphate plate precipitate prepared present protease PROTOCOL provides purification range reaction reagent reducing referred remove residues salt sample sample buffer SDS-PAGE separation sequence soluble solution specific staining standard step structure supernatant surface TABLE target protein technique temperature tion transfer tube usually volume wash Western blotting
Henvisninger til denne bog
Mass Spectrometry of Biological Materials, Second Edition Barbara S. Larsen,Charles N. McEwen Begrænset visning - 1998 |