• формат pdf
  • размер 17.32 МБ
  • добавлен 09 августа 2011 г.
Westermeier R. Electrophoresis in Practice
Wiley-VCH; 3 edition (April 15, 2001)
"The illustrations are informative and to the point.The book clearly illustrates the competence of the author. He generously shares his experience and knowledge with the reader, especially in the field of gel-based electrophoresis.an excellent reference to help the reader to become acquainted with the field of electrophoresis. The book contains theoretical and ample practical information that serves not only the practician in the electrophoresis laboratory, but also students and lecturers in electrophoretic methods."
Contents
Fundamentals
Electrophoresis
General
Electrophoresis in non-restrictive gels
Electrophoresis in restrictive gels
sotachophoresis
Migration with the same speed
on train separation
Zone sharpening effect
Concentration regulation effect
soelectric focusing
Principles
Gels for IEF
Temperature
Controlling the pH gradient
The kinds of pH gradients
Protein detection in IEFgels
Preparative isoelectric focusing
Titration curve analysis
Blotting
Principle
Transfer methods
Blotting membranes
Buffers for electrophoretic transfers
General staining
Blocking
Specific detection
Protein sequencing
Transfer problems
nterpretation of electropherograms
ntroduction
mage analysis
Proteome Analysis
General
Sample preparation
Two-dimensional electrophoresis.
Detection techniques
mage analysis
Protein spot identification
Bioinformatics
Functional proteomics
nstrumentation
Current and voltage conditions
Power supply
Separation chambers
Staining apparatus for gels and blots
Automated electrophoresis
nstruments for 2-D electrophoresis
Safety measures
Environmental aspects
Equipment and Methods
Equipment for Part II
nstrumentation
Special laboratory equipment
Consumables
Chemicals
Method 1: PAGE of dyes
Sample preparation
Stock solutions
Preparing the casting cassette
Casting ultrathin-layer gels
Electrophoretic separation
Method 2: Agarose and immuno electrophoresis
Sample preparation
Stock solutions
Preparing the gels
Electrophoresis
Protein detection
Method 3: Titration curve analysis
Sample preparation
Stock solutions
Preparing the blank gels
Titration curve analysis
Coomassie and silver staining
nterpreting the curves
Method 4: Native PAGE in amphoteric buffers
Sample preparation
Stock solutions
Preparing the empty gels
Electrophoresis
Coomassie and silver staining
Method 5: Agarose IEF
Sample preparation
Preparing the agarose gel
soelectric focusing
Protein detection
Method 6: PAGIEF in rehydrated gels
Sample preparation
Stock solutions
Preparing the blank gels
soelectric focusing
Coomassie and silver staining
Perspectives
Method 7: Horizontal SDS-PAGE
Sample preparation
Stock solutions for gel preparation
Preparing the casting cassette
Gradient gel
Electrophoresis
Protein detection
Blotting
Perspectives
Method 8: Vertical PAGE
Sample preparation
Stock solutions
Single gel casting
Multiple gel casting
Electrophoresis
SDS electrophoresis of small peptides
Two-dimensional electrophoresis
DNA electrophoresis
Long shelflife gels
Detection of bands
Method 9: Semi-dry blotting of proteins
Transfer buffers
Technical procedure
Staining of blotting membranes
Method 10: IEF in immobilized pH gradients
Sample preparation
Stock solutions
mmobiline recipes
Preparing the casting cassette
Preparing the pH gradient gels
soelectric focusing
Staining
Strategies for IPG focusing
Method 11: High-resolution 2-D electrophoresis
Sample preparation
Stock solutions
Preparing the gels
Separation conditions
Staining procedures
Method 12: PAGE of double stranded DNA
Stock solutions
Preparing the gels
Sample preparation
Electrophoresis
Silver staining
Method 13: Native PAGE of single stranded DNA
Sample treatment
Gel properties
Buffers and additives
Conditions for electrophoresis
Strategy for SSCP analysis
Method 14: Denaturing gradient gel electrophoresis
Sample preparation
Rehydration solutions
Preparing the rehydration cassette
Rehydration
Electrophoresis
Silver staining
Method 15: Denaturing PAGE of DNA
Sample preparation
Solutions
Rehydration
Electrophoresis
Silver staining
Appendix Trouble-shooting
soelectric focusing
PAGIEFwith carrier ampholytes
Agarose IEFwith carrier ampholytes
mmobilized pH gradients
SDS electrophoresis
ertical PAGE
Semidry blotting
D electrophoresis
DNA electrophoresis
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