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maintech木蟲 (正式寫手)
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分享-Surface Plasmon Resonance-無重復(fù) 已有1人參與
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1. INTRODUCTION 3 2. PRINCIPLES AND APPLICATIONS OF SURFACE PLASMON RESONANCE. 4 2.1. Principles 4 2.2. Applications 5 2.2.1. What SPR is good for 5 2.2.2. What SPR is not good for. 7 3. GENERAL PRINCIPLES OF BIACORE EXPERIMENTS 8 3.1. A typical experiment 8 3.2. Preparation of materials and buffers 9 3.3. Monitoring the Dips 9 4. LIGAND 10 4.1. Direct versus indirect immobilisation 10 4.2. Covalent immobilisation 11 4.2.1. A general approach 11 4.2.2. Choice of chemistry 12 4.2.3. Prepare the protein 12 4.2.4. Pre-concentration 12 4.2.5. Amine coupling 14 4.2.6. Regeneration 15 4.2.7. Adjusting the immobilisation conditions. 16 4.3. Non-covalent immobilisation (ligand capture) 16 4.3.1. Using an existing strategy 17 4.3.2. Developing a new strategy 17 4.4. Activity of immobilised ligand 18 4.5. Control surfaces 19 4.6. Re-using sensor chips 19 5. ANALYTE 20 5.1. Purity, activity and concentration 20 - 1 - 5.2. Valency 20 5.3. Refractive index effect and control analytes 21 5.4. Low molecular weight analytes 22 6. QUALITITATIVE ANALYSIS; DO THEY INTERACT? 22 6.1. Positive and negative controls 22 6.2. Qualitative comparisons using a multivalent analyte 23 7. QUANTITATIVE MEASUREMENTS 23 7.1. Affinity 24 7.1.1. Concepts 24 7.1.2. Experimental design 24 7.1.3. Data analysis 26 7.1.4. Controls 27 7.1.5. Non-linear Scatchard Plots 27 7.2. Kinetics 30 7.2.1. Concepts 30 7.2.2. Experimental design 31 7.2.3. Data analysis 33 7.2.4. Controls 34 7.3. Stoichiometry 35 7.4. Thermodynamics 35 7.5. Activation energy 37 8. CONCLUSION 38 9. APPENDIX 38 9.1. Physical basis of SPR 38 9.2. Samples and buffers 39 9.2.1. Guidelines for preparing samples and buffers for use on the BIAcore 39 9.2.2. Standard buffers 40 9.3. Additional information 41 10. REFERENCES 41 11. ACKNOWLEDGEMENTS 42 13. FIGURE 1. ANALYSIS OF DIPS. 44 14. TABLES 45 14.1. Table 1. BIAcore instruments currently available (January 1999) 45 14.2. Table 2. Sensor chips available for use on the BIAcore 46 14.3. Table 3. Covalent coupling chemistry 47 14.4. Table 4. Techniques for ligand capture. 48 14.5. Table 5. Trivial causes of complex binding kinetics 49 14.6. Table 6. Distinguishing some non-trivial causes of complex kinetics 50 |
金蟲 (正式寫手)
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