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Rationally Designed Ligands for Use in Affinity Chromatography 95 Fig. 1. Strategy followed for the development of synthetic affinity ligands mimicking the interaction of Protein L with the Fab fragment of immunoglobulins. 2. Materials 2.1. Study of the PpL-Fab Binding Site and De Novo Design of Affinity Ligands 1. Computer-aided molecular modelling: Different software packages are commercially available to perform molecular modelling, including Quanta2000 and InsightII from Accelrys, which can run on a IRIX®6.5 Silicon Graphics®Octane® workstation from Silicon Graphics Inc. Some molecular modelling studies were also carried out in a microsoft windows environment using WebLab<strong>View</strong>erLite (http://www.msi.com), SwissPDB<strong>View</strong>er3.7 (http://www.expasy.ch/spdbv),
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Affinity Chromatography second edit
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METHODS IN MOLECULAR BIOLOGY TM Aff
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To Tina, Emmanuella, Natalie, and A
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viii Preface Affinity tags for puri
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x Contents 10. Immobilized Metal Io
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xii Contributors Tzong-Hsien Lee
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2 Roque and Lowe cell extract conta
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4 Roque and Lowe groups critical in
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6 Roque and Lowe of reinforcement e
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8 Roque and Lowe unknown factors ar
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10 Roque and Lowe receptor domains
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12 Roque and Lowe A further issue o
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14 Roque and Lowe transgenic system
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16 Roque and Lowe 6. Arsenis, C. an
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18 Roque and Lowe 39. Burton, S.J.,
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20 Roque and Lowe 71. Bhikhabhai, R
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I Various Modes of Affinity Chromat
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26 Charlton and Zachariou Since the
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28 Charlton and Zachariou 5. Equili
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30 Charlton and Zachariou 9. Elute
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32 Charlton and Zachariou 3.3. Puri
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34 Charlton and Zachariou could als
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3 Affinity Precipitation of Protein
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Affinity Precipitation of Proteins
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4 Immunoaffinity Chromatography Stu
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Immunoaffinity Chromatography 55 2.
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Immunoaffinity Chromatography 57 A
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Immunoaffinity Chromatography 59 ab
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5 Dye Ligand Chromatography Stuart
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Dye Ligand Chromatography 63 Develo
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Dye Ligand Chromatography 65 6. Mis
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Page 158: 72 Tugcu chromatography, the sorpti
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Page 166: 76 Tugcu the salt counter ions on t
Page 170: 78 Tugcu On a plot of log K versus
Page 174: 80 Tugcu Figure 2B illustrates a ty
Page 178: 82 Tugcu 2.5. Running Displacement
Page 182: 84 Tugcu then the operating conditi
Page 186: 86 Tugcu Table 1 Trobleshooting for
Page 190: 88 Tugcu 23. Torres, A. R. and Pete
Page 194: II Affinity Chromatography Using Pu
Page 198: 94 Roque and Lowe market, with 14 F
Page 204: Rationally Designed Ligands for Use
Page 232: 112 Casey et al. be constructed by
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Page 240: 116 Casey et al. 3. Wash the coated
Page 244: 118 Casey et al. 6) Wash four times
Page 248: 120 Casey et al. 3) Mix the washed
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122 Casey et al. C Absorbance 450nm
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124 Casey et al. References 1. Smit
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126 Forde Utilization of the GST-gl
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128 Forde 3. Methods 3.1. Productio
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130 Forde 4. Wash the column with 5
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132 Forde 5. BDGE is toxic (by inha
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134 Forde 250 Elution in response u
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136 Forde References 1. Wils P, Esc
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138 Charlton and Zachariou 1. Intro
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140 Charlton and Zachariou and non-
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142 Charlton and Zachariou the maxi
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144 Charlton and Zachariou Table 2
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146 Charlton and Zachariou the Tabl
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148 Charlton and Zachariou for some
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11 Methods for the Purification of
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Purification of HQ-Tagged Proteins
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12 Amylose Affinity Chromatography
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Amylose Affinity Chromatography of
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192 Urh et al. and affinity purific
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194 Urh et al. beads with HaloTag l
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196 Urh et al. 2.3. Detection of Pr
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198 Urh et al. 2. Carefully remove
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200 Urh et al. 3.2.1.2. Phase 2 Imm
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202 Urh et al. 3.2.2. Detection of
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204 Urh et al. The following protoc
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206 Urh et al. 3. Incubate for 10 m
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208 Urh et al. by 0.5% Triton X-100
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14 Site-Specific Cleavage of Fusion
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Site-Specific Cleavage of Fusion Pr
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15 The Use of TAGZyme for the Effic
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Removal of N-Terminal His-Tags 231
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16 Affinity Processing of Cell-Cont
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Affinity Processing of Cell-Contain
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258 Benčina et al. 1. Introduction
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260 Benčina et al. 3.1.1. Static I
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262 Benčina et al. [A] abs orbance
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264 Benčina et al. Table 2 Long-Te
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266 Benčina et al. Table 3 Effect
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268 Benčina et al. 8 7 n RNase 0,0
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270 Benčina et al. Table 6 Long-Te
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272 Benčina et al. B A PepC marker
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274 Benčina et al. 3. Platonova, G
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276 Forde diseases, cancer and infe
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278 Forde 12. Sample loading buffer
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280 Forde 2. Mix 100 μl of sample,
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282 Forde 12. Safety Warning: Picog
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19 Affinity Chromatography of Phosp
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Affinity Chromatography of Phosphor
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20 Protein Separation Using Immobil
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Immobilized Phospholipid Chromatogr
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21 Analysis of Proteins in Solution
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Affinity Capillary Electrophoresis
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Index Adsorption isotherm, 75, 84 A
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Index 341 Genenase I, 170, 214, 215
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Index 343 Saccharomyces cerevisae,
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