Gel electrophoresis--proteins : essential techniques /
| Main Author: | |
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| Format: | Book |
| Language: | English |
| Published: |
Chichester ; New York :
John Wiley,
[1996]
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| Series: | Essential techniques series.
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| Subjects: |
Table of Contents:
- Introduction to protein electrophoresis
- Protein electrophoresis
- Sample preparation
- Protein loading
- Protein solubilization
- Solubilization under native conditions
- Solubilization under denaturing conditions
- Estimation of proteins
- Methods available
- The Bradford technique
- The Lowry method
- Bicinchoninic acid
- Amido black filter assay
- Protocols
- Protein estimation by the Bradford technique
- Modified Bradford technique for protein determination in the presence of carrier ampholytes
- Amido black filter assay.
- Gel preparation
- Methods available
- Gel matrices available
- Polyacrylamide gels
- Agarose gels
- Gel casting formats
- Types of gel apparatus
- Electrophoresis in vertical slabs
- Electrophoresis in horizontal slabs
- Electrophoresis in cylindrical gels
- Gradient gels
- Preparation of glassware
- Preparation of backing sheets
- Gel casting for vertical polyacrylamide slab gels
- Gel casting for horizontal polyacrylamide slab gels
- Casting cylindrical polyacrylamide gels
- Agarose slab gel electrophoresis.
- Rehydratable gels
- Preparation of gradient gels using gradient makers
- Preparation of gradients by the tilting method
- Gel techniques
- Methods available
- Separations under native conditions using intrinsic charge
- Tris-glycine gels
- Ferguson analysis
- Electrofocusing
- Focusing with carrier ampholytes in tube gels
- Focusing in horizontal slabs
- Separation under denaturing conditions
- SDS gel electrophoresis
- Two-dimensional electrophoresis
- Special techniques
- Peptide mapping
- Immunoelectrophoresis.
- Rocket immunoelectrophoresis
- Crossed immunoelectrophoresis
- Choice of methods
- Protocols
- Tris-glycine polyacrylamide gels
- Isoelectric focusing in rehydratable polyacrylamide slab gels
- Isoelectric focusing in agarose gels
- Isoelectric focusing in ultrathin gels (pH 5-7)
- Isoelectric focusing using immunobilines
- SDS-polyacrylamide gels
- Two-dimensional electrophoresis with carrier ampholytes using a cylindrical first-dimensional gel and a vertical second-dimensional gel
- Staining for glycoproteins.
- Staining for lipoproteins
- Colloidal coomassie blue staining
- Silver staining of gels
- Radiofluorography
- Identification of proteins in gels
- Elution and renaturation
- Methods available
- Immunochemical identification
- Western blotting
- Immunosubtraction
- Associated methodologies
- Zymographic identification
- Infusion of substrate
- Overlay matrices
- Activity gels
- Choice of methods
- Protocols
- Capillary blotting from a polyacrylamide slab gel to nitrocellulose
- Vertical electroblotting to nitrocellulose nylon-66 or PVDF.
- Northwestern blotting
- Immunosubstraction
- Infusion of reagents into gels-demonstration of †-galactosidase
- One-step zymogram using an indicator matrix-demonstration of DNase
- Coupled zymogram using an indicator matrix-demonstration of phosphoglucomutase
- Activity gels
- Recording and storing gels
- Methods available
- Computerized methods
- Unidimensional scanning
- Photocopying, photographing and duplicating
- Drying
- Protocols
- Recording gel patterns using the electrophoresis duplicating system
- Photographing gels.