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Index 343 Saccharomyces cerevisae, 295 Scatchard plot equation, 107 Screening techniques, 97 secondary, 69 Secreted HQ-Tagged proteins, 161 Sequencing of clones, 118 SMA isotherm, 75–77, 79–80, 84 Soham Scientific Ltd, 135 Solid-phase assembly, 4–5 combinatorial chemistry, 14, 93 combinatorial synthesis, 100 synthesis of lead ligands, 101 Spin columns centrifugation protocol for, 156 vacuum protocol for, 157 “Square-wave” zones, 72 Static immobilization method, 260 Stationary phase, identification of, 75 STREP tag, 37 Synthetic peptide, characterization of the, 119 Tag removal step, 230 TAGZyme, 229–235, 238 Thioredoxin, 13, 38, 139 Three-dimensional matrix environment, 8 Thrombin, 213–214, 217 Tiselius, 72 Titration of phage, 118 TNT ® quick coupled transcription, 162 Tobacco Etch Virus (TEV), 215 “Traditional” pseudobiospecific affinity matrices, 94 Trypan blue dye exclusion method, 254 Trypsin enzyme, 269–270 Trypsin immobilization, 268 Tumor necrosis factor, 235 Two-dimensional electrophoresis (2D-PAGE), 13, 291, 301 U.S. Patent Office, 139 Ultrafiltration, 68, 84 Unclarified lysate, 129 Viral cysteine proteases, 215 Wheat germ extract, 153 X-ray crystallographic structures, 14 Yeast tryptone (YT) media, 115 Zinc finger transcription factor, 125–126, 275
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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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Dye Ligand Chromatography 67 Fig. 1
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Dye Ligand Chromatography 69 6. Sec
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72 Tugcu chromatography, the sorpti
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74 Tugcu non-specific interactions,
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76 Tugcu the salt counter ions on t
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78 Tugcu On a plot of log K versus
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80 Tugcu Figure 2B illustrates a ty
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82 Tugcu 2.5. Running Displacement
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84 Tugcu then the operating conditi
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86 Tugcu Table 1 Trobleshooting for
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88 Tugcu 23. Torres, A. R. and Pete
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II Affinity Chromatography Using Pu
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94 Roque and Lowe market, with 14 F
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96 Roque and Lowe and RasMol V2.7.1
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98 Roque and Lowe house using, for
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100 Roque and Lowe Gly71 and Gly72)
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102 Roque and Lowe dissolved in ace
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104 Roque and Lowe equilibration bu
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106 Roque and Lowe of color. Purple
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108 Roque and Lowe 5. Fassina, G.,
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8 Phage Display of Peptides in Liga
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Phage Display of Peptides in Ligand
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9 Preparation, Analysis and Use of
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Preparation, Analysis and Use of an
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10 Immobilized Metal Ion Affinity C
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IMAC of Histidine-Tagged Fusion Pro
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152 Godat et al. tag. HQ-tagged pro
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154 Godat et al. 2. NaCl (5 M). 3.
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156 Godat et al. 4. Invert tube to
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158 Godat et al. 3. Sonicate sample
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160 Godat et al. the above protocol
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162 Godat et al. 3.7.1. Purificatio
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164 Godat et al. 3. Adding 200 mM N
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166 Godat et al. 4. The MagneHis Ni
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168 Godat et al. 22. Lin, D., Tabb,
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170 Pattenden and Thomas 1. Introdu
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172 Pattenden and Thomas functions
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174 Pattenden and Thomas of the mal
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176 Pattenden and Thomas necessary.
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178 Pattenden and Thomas 2.4. Amylo
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180 Pattenden and Thomas 9. Thoroug
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182 Pattenden and Thomas 8. Collect
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184 Pattenden and Thomas binding ef
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186 Pattenden and Thomas 15. Cells
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188 Pattenden and Thomas 23. Martin
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13 Methods for Detection of Protein
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Detection of Protein-Protein and Pr
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212 Charlton recognition sequence,
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214 Charlton when selecting Factor
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216 Charlton 2. Materials 2.1. Reag
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218 Charlton (see Notes 12 and 13).
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220 Charlton 3.3. Cleavage of Fusio
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222 Charlton 3. The catalytic mecha
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224 Charlton may dictate. However,
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226 Charlton 22. The full-length fu
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228 Charlton 22. Lien, S., Milner,
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230 Arnau et al. downstream process
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232 Arnau et al. Fig. 1. Overview o
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234 Arnau et al. Fig. 2. The pQE-1
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236 Arnau et al. 2.3.3. Step B Buff
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238 Arnau et al. Fig. 3. Immobilize
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240 Arnau et al. 6. Collect the flo
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242 Arnau et al. 4. Desalting is ne
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III Various Applications of Affinit
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248 Galaev and Mattiasson in biotec
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250 Galaev and Mattiasson 2.4. Sepa
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252 Galaev and Mattiasson 3.4. Sepa
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254 Galaev and Mattiasson ensure el
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17 Monolithic Bioreactors for Macro
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Monolithic Bioreactors for Macromol
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18 Plasmid DNA Purification Via the
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Plasmid DNA Purification 277 are pr
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Plasmid DNA Purification 279 5. Dec
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Plasmid DNA Purification 281 Initia
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Plasmid DNA Purification 283 8. Bou
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286 Tchaga proteins is variable. Ho
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288 Tchaga residue is buried and is
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290 Tchaga 3. Centrifuge the tube a
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292 Tchaga 5. Use the BCA Protein A
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294 Tchaga 24. Figeys D., Gygi S.P.
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296 Lee and Aguilar The primary dif
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298 Lee and Aguilar 2.2. Equipment
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300 Lee and Aguilar Abs 280 PLA 2 A
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302 Lee and Aguilar 9. Pidgeon, C.,
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304 Heegaard et al. 1. Introduction
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Table 1 CE Methods and Their Corres
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308 Heegaard et al. Fig. 1. Images
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310 Heegaard et al. characteristics
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312 Heegaard et al. sample. A final
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314 Heegaard et al. Especially usef
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316 Heegaard et al. parameters extr
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Page 658: 332 Heegaard et al. 21. Grossman, P
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Page 666: 336 Heegaard et al. 82. Berezovski,
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Page 674: 340 Index Cibacron blue, 6-7, 61 Cl
Page 678: 342 Index Monogenic diseases, 275 M
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