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A Laboratory Guide to Biotin-Labeling in Biomolecule Analysis (Biomethods) - Softcover

 
9783034873512: A Laboratory Guide to Biotin-Labeling in Biomolecule Analysis (Biomethods)

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2.1 Introduction ........................................................... 32 2.2 Technical Procedures ................................................... 32 Chemical Synthesis ..................................................... 35 2.3 Results and Discussion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . 38 . . . . . . . . . . . . . Chemical Synthesis ..................................................... 38 Binding to anti-CCK antisera and CCK-B receptors .......................... 40 Binding to streptavidin agarose and photoelution ........................... 40 Affinity chromatography ................................................. 41 2.4 Troubleshooting....................................................... 43 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . 44 . . . . . . . . . . . . . . . . . Purification of the Receptor for Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) using Biotinylated Ligands ......................... 45 Summary ............................................................. 45 3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . 46 . . . . . . . . . . . . . . . . . 3.2 Technical Procedures ................................................... 48 Preparation of a Biotinylated Ligand ...................................... 48 Preparation of Membrane Fraction ........................................ 50 Solubilization of the PACAP Receptor ..................................... 51 Partial Purification of the PACAP Receptor ................................. 52 Affinity Purification of the PACAP Receptor ................................ 54 Final Purification of the PACAP Receptor .................................. 56 Receptor-Binding Assay ................................................. 57 3.3 Results and Discussion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . 58 . . . . . . . . . . . . . 3.4 Troubleshooting ....................................................... 61 Impurity in PACAP27-Cys-NH............................................ 61 2 Low specific activity of the PACAP receptor despite a single band ............. 62 Acknowledgments ..................................................... 62 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . 63 . . . . . . . . . . . . . . . . . Photoreactive Biotinylated Peptide Ligands for Affinity Labeling ......... " 65 Summary ............................................................. 65 4.1 Introduction................................................ . . . . . . . .. . 66 . 4.2 Technical Procedures ................................................. " 67 Synthesis of a trifunctional photoactivatable biotinylating reagent ............. 67 Synthesis of photo reactive biotinylated peptide hormones ................... 67 Site-specific incorporation of biotin and photo labels in separate steps ........ 69 Photoaffinity labeling ................................................... 72 4.3 Results and Discussion .................................................. 76 Synthesis of photoactivatable insulins with permanent biotin labels ............ 76 Applications: Insulin .................................................... 78 VI Examples for other applications .......................................... 78 4.4 Troubleshooting....................................................... 79 References . . . . . . . . . . . . . . . . . . . . .

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  • PublisherBirkhäuser
  • Publication date2012
  • ISBN 10 3034873514
  • ISBN 13 9783034873512
  • BindingPaperback
  • LanguageEnglish
  • Number of pages254
  • EditorMeier T., Fahrenholz F.

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Taschenbuch. Condition: Neu. This item is printed on demand - it takes 3-4 days longer - Neuware -2.1 Introduction . 32 2.2 Technical Procedures . 32 Chemical Synthesis . 35 2.3 Results and Discussion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 . . . . . . . . . . . . . Chemical Synthesis . 38 Binding to anti-CCK antisera and CCK-B receptors . 40 Binding to streptavidin agarose and photoelution . 40 Affinity chromatography . 41 2.4 Troubleshooting. 43 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 . . . . . . . . . . . . . . . . . Purification of the Receptor for Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) using Biotinylated Ligands . 45 Summary . 45 3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 . . . . . . . . . . . . . . . . . 3.2 Technical Procedures . 48 Preparation of a Biotinylated Ligand . 48 Preparation of Membrane Fraction . 50 Solubilization of the PACAP Receptor . 51 Partial Purification of the PACAP Receptor . 52 Affinity Purification of the PACAP Receptor . 54 Final Purification of the PACAP Receptor . 56 Receptor-Binding Assay . 57 3.3 Results and Discussion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58 . . . . . . . . . . . . . 3.4 Troubleshooting . 61 Impurity in PACAP27-Cys-NH. 61 2 Low specific activity of the PACAP receptor despite a single band . 62 Acknowledgments . 62 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 . . . . . . . . . . . . . . . . . Photoreactive Biotinylated Peptide Ligands for Affinity Labeling . ' 65 Summary . 65 4.1 Introduction. . . . . . . . . . 66 . 4.2 Technical Procedures . ' 67 Synthesis of a trifunctional photoactivatable biotinylating reagent . 67 Synthesis of photo reactive biotinylated peptide hormones . 67 Site-specific incorporation of biotin and photo labels in separate steps . 69 Photoaffinity labeling . 72 4.3 Results and Discussion . 76 Synthesis of photoactivatable insulins with permanent biotin labels . 76 Applications: Insulin . 78 VI Examples for other applications . 78 4.4 Troubleshooting. 79 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81 . . . . . . . . . . . . . . . . . . . 240 pp. Englisch. Seller Inventory # 9783034873512

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Taschenbuch. Condition: Neu. Druck auf Anfrage Neuware - Printed after ordering - 2.1 Introduction . 32 2.2 Technical Procedures . 32 Chemical Synthesis . 35 2.3 Results and Discussion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 . . . . . . . . . . . . . Chemical Synthesis . 38 Binding to anti-CCK antisera and CCK-B receptors . 40 Binding to streptavidin agarose and photoelution . 40 Affinity chromatography . 41 2.4 Troubleshooting. 43 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 . . . . . . . . . . . . . . . . . Purification of the Receptor for Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) using Biotinylated Ligands . 45 Summary . 45 3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 . . . . . . . . . . . . . . . . . 3.2 Technical Procedures . 48 Preparation of a Biotinylated Ligand . 48 Preparation of Membrane Fraction . 50 Solubilization of the PACAP Receptor . 51 Partial Purification of the PACAP Receptor . 52 Affinity Purification of the PACAP Receptor . 54 Final Purification of the PACAP Receptor . 56 Receptor-Binding Assay . 57 3.3 Results and Discussion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58 . . . . . . . . . . . . . 3.4 Troubleshooting . 61 Impurity in PACAP27-Cys-NH. 61 2 Low specific activity of the PACAP receptor despite a single band . 62 Acknowledgments . 62 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 . . . . . . . . . . . . . . . . . Photoreactive Biotinylated Peptide Ligands for Affinity Labeling . ' 65 Summary . 65 4.1 Introduction. . . . . . . . . . 66 . 4.2 Technical Procedures . ' 67 Synthesis of a trifunctional photoactivatable biotinylating reagent . 67 Synthesis of photo reactive biotinylated peptide hormones . 67 Site-specific incorporation of biotin and photo labels in separate steps . 69 Photoaffinity labeling . 72 4.3 Results and Discussion . 76 Synthesis of photoactivatable insulins with permanent biotin labels . 76 Applications: Insulin . 78 VI Examples for other applications . 78 4.4 Troubleshooting. 79 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81 . . . . . . . . . . . . . . . . . . . Seller Inventory # 9783034873512

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Condition: New. Dieser Artikel ist ein Print on Demand Artikel und wird nach Ihrer Bestellung fuer Sie gedruckt. 1 An Introduction to Avidin-Biotin Technology and Options for Biotinylation.- 1.1 Introduction.- The Avidin-Biotin Interaction.- Avidin.- Streptavidin.- Biotin.- 1.2 Technical Procedures.- Determining Biotin and Biotin Incorporation Levels.- Biotinylation R. Seller Inventory # 4319092

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