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Table Of Contents

1.1 Principles of Biacore technology
1.2 Biacore terminology
1.3.1 The SPR detection system
1.3.2 The sensor surface
1.3.3 The microfluidic system
1.4 Scope of this handbook
1.5 Additional information
2.1.1 The gold layer
2.1.2 The surface coating
2.2.1 CM-series sensor chips
2.2.2 Sensor chips for capture applications
2.2.3 Sensor chips for hydrophobic ligands
2.2.4 Sensor chips for special applications
3.1 Covalent immobilization
3.2.1 Streptavidin-biotin capture
3.2.2 Antibody-based capture
3.2.3 Capturing tagged recombinant proteins
3.3.1 Monolayer attachment
3.3.2 Bilayer attachment
3.4 Immobilizing small molecules
3.5 Techniques for stabilizing the surface
3.6.1 How much ligand should be immobilized?
3.6.2 Choice of sensor surface
3.6.3 Choice of immobilization method
4.1.1 Temperature
4.1.2 Buffer pH and ligand pre-concentration
4.1.3 Other buffer conditions
4.1.4 Ligand concentration
4.1.5 Method for determining coupling conditions
4.2.1 Preparing solutions
4.2.2 Evaluating covalent immobilization
4.2.3 Testing the surface after immobilization
4.3 Activating the surface
4.4.2 Required solutions
4.4.3 Immobilization procedure
4.5.1 Ligand thiol coupling
4.5.2 Surface thiol coupling
4.5.3 Maleimide coupling
4.6.1 Chemistry
4.6.2 Oxidizing the ligand
4.6.3 Required solutions
4.6.4 Immobilization procedure
4.6.5 Results
4.7.1 Biotinylation for streptavidin-biotin capture
5.1.1 Preparation for use
5.1.2 Liposome preparation and adsorption
5.2.1 Preparations for use
5.2.2 Liposome preparation and adsorption
5.3 Liposome attachment on other sensor chips
5.4.1 Preparation of ligand and micelles
5.4.2 Immobilizing the ligand
5.4.3 Lipid deposition and membrane reconstitution
6.1.1 General considerations
6.1.2 Choice of regeneration solution
6.1.3 Factors affecting regeneration
6.1.4 Regeneration scouting procedure
6.2.1 Interpretation principles
6.2.2 Case studies
6.3 Verifying regeneration conditions
6.4.1 Protein surfaces
6.4.2 Nucleic acids
6.4.3 Membrane-associated ligands
6.4.4 Small molecules
6.4.5 Other regeneration strategies
7.1 Testing the surface
7.2.1 Low immobilization levels
7.2.2 Low analyte responses
7.2.3 When regeneration fails
7.2.4 Problems with non-specific binding
8.1 Recommended procedures – wet storage
8.2 Recommended procedures – dry storage
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Sensor Surface Handbook

Sensor Surface Handbook

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Published by mc2camila

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Published by: mc2camila on May 05, 2011
Copyright:Attribution Non-commercial

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12/17/2012

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