Western blotting guru / Ayaz Najafov, Gerta Hoxhaj.
Material type:
TextPublication details: London, U.K. : Academic Press is an imprint of Elsevier, [2017]Description: 1 online resourceContent type: text Media type: computer Carrier type: online resourceISBN: 0128135387; 9780128135389Subject(s): Western immunoblotting -- Laboratory manuals | Molecular biology | SCIENCE -- Life Sciences -- Biochemistry | Molecular biology | Western immunoblotting | Blotting, Western -- methodsGenre/Form: Electronic books. | Laboratory manuals. | Electronic book.DDC classification: 572/.636 LOC classification: QP519.9.W47NLM classification: 2017 G-792 | QW 525.5.I32Online resources: ScienceDirect | ScienceDirect | Item type | Current library | Call number | Status | Date due | Barcode |
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Ebooks
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Mysore University Main Library | Not for loan | EBKELV1008 |
Includes bibliographical references.
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.
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 in molecular biology laboratories. The book covers the basic science underlying immunoblotting and detailed description of the method parameters, followed by good benchtop practices, tips and tricks for obtaining high-quality data and a detailed troubleshooting guide addressing a variety of problem types.
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