2.4 Cross-linking of Ligands
2.4.1 Three-component Cross-linking (3C Cross-linking)
2.4.2 Photofinity Cross-linking
2.4.3 Controls for Cross-linking Experiments
2.5 Purposes
3 Solubilization of Membrane Proteins
3.1 Detergents
3.1.1 Clean Concepts
3.1.2 Handling Detergents
3.2 Solubilization
3.2.1 Solubilization Criteria
3.2.2 Physical Parameters of Solubilized Membrane Proteins
4 Protein Detection via Functional Measurements
4.1 Translocators
4.1.1 Liposomes
4.1.2 Proteoliposomes
4.2 Reconstitution
4.2.1 Reconstitution from a Solution
4.2.2 Reconstitution in Preformed Liposomes
4.3 Flux Assay
4.3.1 Influx Assay
4.3.2 Efflux Assay
4.4 Constructive Thoughts
5 Cleaning and Purifying
5.1 Pure Fun
5.2 Conventional Purification Methods
5.2.1 The Column Technique
5.2.2 Purification Based on Size Differences
5.2.3 Purification Based on Charge Differences
5.2.4 Hydrophobic Chromatography
5.2.5 The Blue Gel
5.3 Affinity Chromatography
5.3.1 Lectin Chromatography
5.3.2 Ligand Chromatography
5.4 The Purity Test
5.5 Profiting
6 Antibodies
6.1 Production of Polyclonal Antibodies
6.1.1 Antigen
6.1.2 Adjuvant
6.1.3 Injection and Serum Harvesting
6.1.4 Purification of Antibodies
6.2 Immunoprecipitation
6.2.1 Immunoprecipitation with Immobilized Protein A
6.2.2 Immunoprecipitation with Immobilized Antibody
6.3 Immunoaffinity Chromatography
6.4 Antibodies Against Unpurified Proteins
6.5 Immunological Detection Techniques
7 Proteomics
7.1 Introduction