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BL
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851755
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| Title & Author
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Collagen : : methods and protocols /\ edited by Irit Sagi and Nikolaos A. Afratis.
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| Publication Statement
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New York, NY :: Humana Press :: Springer,, [2019]
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, ©2019
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| Series Statement
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Methods in molecular biology,; 1944
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| Page. NO
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1 online resource (xi, 254 pages) :: illustrations (some color)
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| ISBN
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1493990950
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: 9781493990955
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9781493990948
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| Bibliographies/Indexes
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Includes bibliographical references and index.
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| Contents
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Intro; Preface; Contents; Contributors; Part I: In Vitro Models for Collagen Characterization; Chapter 1: Cloning and Mutagenesis Strategies for Large Collagens; 1 Introduction; 2 Materials; 2.1 Polymerase Chain Reaction (PCR); 2.2 Gibson Reaction and Bacterial Transformation; 2.3 Recombinant Expression of Collagen VII; 3 Methods; 3.1 Strategy Design and Generation of DNA Fragments with Overlapping Ends; 3.2 Gibson Assembly and Bacterial Transformation; 3.3 Collagen VII Expression in HEK293 Cells; 3.4 First Assessment of Functionality of Engineered Collagen VII Variants; 4 Notes; References
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2.6 Metabolic Activity Assay (alamarBlue)2.7 Cell Viability Assay; 3 Methods; 3.1 Collagen Cross-Linking and Film Preparation; 3.2 Swelling Assay; 3.3 Tensile Test; 3.4 Cell Culture; 3.4.1 Scaffold Preparation; 3.4.2 DF Seeding; 3.4.3 THP-1 Seeding; 3.5 Cell Proliferation and Morphology Assay; 3.6 Cell Metabolic Activity Assay (alamarBlue); 3.7 Cell Viability Assay; 4 Notes; References; Chapter 4: A Biomimetic Model for Mineralization of Type-I Collagen Fibrils; 1 Introduction; 2 Materials; 3 Methods; 3.1 Conventional TEM Experiments; 3.1.1 Self-Assembly of Type-I Collagen Fibrils
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3.1.2 Collagen Mineralization3.1.3 TEM Imaging and Mineralization Degree Calculation; 3.2 CryoTEM Experiments; 3.2.1 Self-Assembly of Type-I Collagen Extracted from Rat Tail Tendon; 3.2.2 Self-Assembly of Type-I Collagen Extracted from Horse Tendon; 3.2.3 Collagen Mineralization; 3.2.4 CryoTEM Imaging; 3.3 STORM Experiments; 3.3.1 Modification of the Laser Confocal Culture Dish (LCCD) and Self-Assembly of Type-I Collagen Fibrils; 3.3.2 Staining and Mineralization of the Type-I Collagen Fibrils; 3.3.3 STORM Imaging; 4 Notes; References; Part II: Proteomics Analysis of Collagen
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Chapter 2: Multi-well Fibrillation Assay as a Tool for Analyzing Crosslinking and Stabilization of Collagen1 Introduction; 2 Materials; 2.1 Collagen Isolation; 2.2 Collagen Fibrillation; 2.2.1 Fibrillation Buffer; 3 Methods; 3.1 Collagen Isolation; 3.2 Collagen Fibrillation; 4 Notes; References; Chapter 3: Production and Characterization of Chemically Cross-Linked Collagen Scaffolds; 1 Introduction; 2 Materials; 2.1 Collagen Cross-Linking and Film Preparation; 2.2 Swelling Assay; 2.3 Tensile Test; 2.4 Cell Culture; 2.5 Cell Proliferation and Morphology Assay
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Chapter 5: Investigation of Triple-Helix Collagen Hydroxylation by Solid-State NMR Spectroscopy1 Introduction; 1.1 NMR Overview; 1.2 Chemical Shift; 1.3 Solid-State NMR and Magic Angle Spinning; 1.4 Sensitivity of NMR Experiments; 1.4.1 Isotopic Enrichment; 1.4.2 Dynamic Nuclear Polarization in Solid-State NMR; 2 Materials; 2.1 NMR Equipment; 2.2 Solid Biological Samples; 2.3 Stable Radicals for DNP Enhancement; 3 Methods; 3.1 Preparing the Radical Stock Solution for DNP Enhancement; 3.2 Packing the ssNMR Rotor; 3.3 Inserting Rotor into Magnet and Magic Angle Spinning
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| Subject
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Collagen, Laboratory manuals.
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Procollagen N-Endopeptidase.
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Collagen.
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Collagen.
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Histological Techniques-- methods.
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| Dewey Classification
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572/.67
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| LC Classification
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QP552.C6
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| Added Entry
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Afratis. Nikolaos A.
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Sagi, Irit
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