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Western blotting guru /
Western Blotting Guru provides researchers in molecular biology with a handy reference for approaching and solving challenging problems associated with immunoblotting setup and optimization. As a laboratory guide, it emphasizes the technical aspects of efficiently employing immunoblotting as a tool...
| Clasificación: | Libro Electrónico |
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| Autor principal: | |
| Otros Autores: | |
| Formato: | Electrónico eBook |
| Idioma: | Inglés |
| Publicado: |
London, U.K. :
Academic Press is an imprint of Elsevier,
[2017]
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| Temas: | |
| Acceso en línea: | Texto completo |
Tabla de Contenidos:
- Front Cover; Western Blotting Guru; Copyright Page; Contents; Preface; Note to the Reader; 1 Introduction; 1.1 What Is Western Blotting?; 1.2 A Bit of History; 2 Procedure; 2.1 Sample Preparation; 2.2 Gel Electrophoresis; 2.2.1 Gel Selection and Preparation; 2.2.2 Running the Gels; 2.2.3 Molecular Weight Markers; 2.3 Protein Transfer; 2.3.1 Transfer Setup; 2.3.2 Choosing the Right Membrane; 2.3.3 Blotting Paper; 2.3.4 Transfer Buffer; 2.3.5 Transfer Power Settings; 2.3.6 Visualization of Proteins After Transfer; 2.4 Blocking the Membrane; 2.5 Primary Antibodies; 2.6 Secondary Antibodies.
- 2.7 Blot Washes2.8 Developing Western Blots; 3 Good Practices; 4 Optimization and Troubleshooting; 4.1 Optimization Rules; 4.2 General Optimization Strategies; 4.2.1 Antibody Concentration; 4.2.2 Antibody Incubation Times; 4.2.3 Wash Stringency; 4.2.4 Blocking; 4.2.5 Preclear the Antibodies; 4.2.6 Supplement Antibody Solutions With Nonphosphorylated Peptides; 4.3 Troubleshooting Specific Problems; 4.3.1 Problem Type 1; 4.3.1.1 Subtype 1; 4.3.1.2 Subtype 2; 4.3.2 Problem Type 2; 4.3.3 Problem Type 3; 4.3.4 Problem Type 4; 4.3.5 Problem Type 5; 4.3.6 Problem Type 6; 4.3.7 Problem Type 7.
- 4.3.8 Problem Type 84.3.9 Problem Type 9; 4.3.10 Problem Type 10; 5 Tips and Tricks; 6 Special Cases; 6.1 Quantitative Western Blotting; 6.2 Overlay Assays; 6.3 Phospho-Specific Antibodies; 6.4 Phos-Tag; 6.5 Nonreducing PAGE; 6.6 Dot Blots; 7 Data Analysis, Storage, Retrieval; Appendix A: Buffers and Solutions; Appendix B: SDS-PAGE Gel Tables; Appendix C: SDS-PAGE Protocol; Appendix D: Wet Transfer and Immunoblotting Protocol; Appendix E: Home-Made Enhanced ChemiLuminescence (ECL) Detection; Appendix F: Stripping Protocols; Appendix G: Coomassie Staining Protocol.
- Appendix H: Lysis of Cells Using Native ConditionsAppendix I: Quick Denaturing Lysis Protocol; Appendix J: Protein Tags; Appendix K: Covalent Crosslinking of Antibodies to Beads; References; Back Cover.