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Record Nr. |
UNINA9910961783603321 |
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Autore |
Wenk Markus R |
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Titolo |
A manual for biochemistry protocols / / Markus R. Wenk, Aaron Zefrin Fernandis |
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Pubbl/distr/stampa |
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Singapore ; ; Hackensack, NJ, : World Scientific, c2007 |
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ISBN |
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9786611121150 |
9781281121158 |
1281121150 |
9789812708311 |
9812708316 |
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Edizione |
[1st ed.] |
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Descrizione fisica |
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Collana |
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Manuals in biomedical research ; ; v. 3 |
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Altri autori (Persone) |
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Disciplina |
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Soggetti |
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Lingua di pubblicazione |
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Formato |
Materiale a stampa |
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Livello bibliografico |
Monografia |
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Note generali |
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Nota di bibliografia |
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Includes bibliographical references and index. |
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Nota di contenuto |
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Intro -- Contents -- Preface -- List of Figures -- List of Contributors -- A Protein Purification -- Summary -- A.1 Protein Precipitation -- A.2 Column Chromatography -- A.2.1 Ionic Exchange Chromatography -- A.2.2 Size-Exclusion Chromatography -- A.2.3 Affinity Chromatography -- A.2.4 Purification of Recombinant Proteins -- A.2.5 Commercially Prepacked Column Kits -- Notes -- B Protein Analysis -- B.1 Protein Estimation -- B.1.1 Absorbance Assays -- B.1.2 Colourimetric Assays -- B.1.3 Commercial Protein Estimation Kits -- B.2 Spectrometric Analysis -- B.3 SDS-PAGE -- B.4 Gel Staining -- B.4.1 Coomassie Blue Staining -- B.4.2 Silver Staining -- B.5 Western Blotting -- B.6 Immunoprecipitations -- Notes -- C Lipid Analysis -- C.1 Lipid Extraction -- C.1.1 Modified Bligh and Dyers Method for Phospholipid Extraction -- C.1.2 Folch Extraction -- C.1.3 Grey Method for Phosphatidylinositol Phosphate Extraction -- C.1.4 Modified Alex Brown Method for Phosphatidylinositol Phosphate Extraction -- C.1.5 Hexane Extraction for Neutral Lipids -- C.1.6 Glycolipid Extraction -- C.2 Thin Layer Chromatography -- Notes -- D Mammalian Cell Culture -- D.1 Primary Culture -- D.1.1 Primary Cell Isolation and Culturing -- |
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D.1.2 Splenocyte Isolation -- D.1.3 Isolation of Peripheral Blood Lymphocytes -- D.2 Tissue Culture -- D.2.1 Adherent Cells -- D.2.2 Suspension Cells -- D.2.3 Cell Maintenance -- D.3 Cell Count -- D.4 Mammalian Transfection -- D.4.1 Calcium Phosphate Coprecipitation -- D.4.2 Electroporation Protocol -- D.4.3 Lipofectamine Transfection Protocol -- Notes -- E Microscopy -- E.1 Poly-L-Lysine Immobilisation -- E.2 Immunofluorescence and Confocal Microscopy -- E.3 Immunohistochemistry -- Notes -- F Assays -- F.1 Enzyme Assays -- F.1.1 Enzyme Kinetics: Basics -- F.1.2 Lipid Kinase Assay -- F.1.3 Protein Kinase Assay. |
F.1.4 Protein Tyrosine Phosphatase Assay -- F.1.5 Alkaline Phosphatase Assay -- F.1.6 Caspase Assay -- F.2 Functional Assays -- F.2.1 Apoptosis Assay -- F.2.2 XTT Cell Proliferation Assay [(2,3-Bis (2-methoxy-4-nitro-5-sulfophenyl)- 2H-tetrazolium-5-carboxanilide)] -- F.2.3 Chemotaxis Assay -- F.2.4 Matrigel Invasion or Chemoinvasion Assay -- Notes -- G Autoradiography -- Summary -- Notes -- Suggested Readings -- Abbreviations -- Appendices -- Appendix 1 Safety Issues -- Appendix 2 Buffer Chart -- Appendix 3 Composition of Regular Buffers or Stock Solutions -- Index. |
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Sommario/riassunto |
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Biochemistry plays an important role in all areas of the biological and medical sciences. With most of the research or diagnosis involved in these areas being based on biochemically obtained observations, it is essential to have a profile of well standardized protocols. This manual is a basic guide for all students, researchers and experts in biochemistry, designed to help readers in directly starting off their experiments without prior knowledge of the protocol. The book dwells on the concepts used in designing the methodologies, thereby giving ample room for researchers to modify them according to their research requirements. |
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