Protein Protocols Protein Protocols

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xxii Contributors SERGEY M. KIPRIYANOV • Affimed Therapeutics AG, Ladenburg, Germany CHRISTIAN KLEIST • Institute for Immunology, Heidelberg, Germany JOACHIM KLOSE • Institut für Humangenetik Charité, Humboldt-Universität, Berlin, Germany. SUNIL KOCHHAR • Nestlé Research Center, Lausanne, Switzerland NICHOLAS J. KRUGER • Department of Plant Sciences, University of Oxford, UK JUDITH A. LAFFIN • Department of Microbiology, Immunology, and Molecular Genetics, The Albany Medical College, Albany, NY WILLIAM J. LAROCHELLE • Laboratory of Cellular and Molecular Biology, National Cancer Institute, National Institute of Health, Bethesda, MD ROBERT R. LATEK • Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, MO JOHN M. LEHMAN • Center for Immunology and Microbial Disease, Albany Medical College, NY MICHELE LEARMONTH • Department of Biomedical Sciences, University of Edinburgh, Scotland ANDRÉ LE BIVIC • IBDM, Faculté des Sciences de Luminy, Marseille, France YEAN KIT LEE • Division of Medical Oncology, Mayo Clinic, Rochester, MN PETER LEMKIN • LECB/NCI-FCRDC, Frederick, MD KRISTI A. LEWIS • Laboratory of Cellular and Molecular Biology, National Cancer Institute, National Institute of Health, Bethesda, MD SABRINA LIBERATORI • Department of Molecular Biology, University of Siena, Italy FAN LIN • Department of Pathology, Temple University Hospital, Philadelphia, PA Mary F. Lopez • Proteome Systems, Woburn, MA BARBARA MAGI • Department of Molecular Biology, University of Siena, Italy NGUYEN THI MAN • MRIC, North East Wales Institute, Deeside, Clwyd, UK SU-YAU MAO • Department of Immunology and Molecular Genetics, Medimmune Inc., Gaithersburg, MD PHILIP N. MCFADDEN • Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR PAUL MCGEADY • Department of Chemistry, Clark Atlanta University, Georgia TONY MERRY • Department of Biochemistry, The Glycobiology Institute, University of Oxford, UK GRAEME MILLIGAN • Department of Biochemistry & Molecular Biology, Institute of Biomedical and Life Sciences, University of Glasgow, UK JONATHAN MINDEN • Millennium Pharmaceuticals, Cambridge, MA THIERRY MINI • Department of Biochemistry, Biozentrum der Universität Basel, Switzerland SHEENAH M. MISCHE • <strong>Protein</strong>/DNA Technology Center, Rockefeller University, NY HOLGER J. MØLLER • Department of Clinical Biochemistry, Aarhus University Hospital, Amtssygehuset, Aarhus, Denmark GLENN E. MORRIS • MRIC, North East Wales Institute, Wrexhäm, UK BARBARA MOURATOU • Biochemisches Institut der Universität Zürich, Switzerland DANIEL MOYNET• INSERM, Bordeaux Cedex, France HIKARU NAGAHARA • Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, MO
Contributors xxiii STEFANIE A. NELSON • Laboratory of Cellular and Molecular Biology, National Cancer Institute, National Institute of Health, Bethesda, MD TOSHIAKI OSAWA • Yakult Central Institute for Microbiology Research, Tokyo, Japan NICOLLE PACKER • Proteome Systems Ltd., Sydney, Australia MARK PAGE • Apovia Inc., San Diego, CA VITALIANO PALLINI • Department of Molecular Biology, University of Siena, Italy GIOVANNA PALOMBO • Biopharmaceuticals, Tecnogen SCPA, Parco Scientifico, Piana di Monte Verna (CE), Italy SCOTT D. PATTERSON • Celera Genomics, Rockville, MD WAYNE F. PATTON • Molecular Probes Inc., Eugene, OR JERGEN H. POULSEN • Department of Clinical Biochemistry, Aarhus University Hospital, Amtssygehuset, Aarhus, Denmark THIERRY RABILLOUD • DBMS/BECP, CEA-Grenoble, Grenoble, France ROBERTO RAGGIASCHI • Department of Molecular Biology, University of Siena, Italy MENOTTI RUVO • Biopharmaceuticals, Tecnogen SCPA, Parco Scientifico, Piana di Monte Verna (CE), Italy F. ANDREW RAY • Department of Biology, Hartwick College, Oneonta, NY JEFFREY ROHRER • Dionex Corporation, Life Science Research Group, Sunnyvale, CA DOUGLAS D. ROOT • Department of Biological Sciences, University of North Texas, Denton, TX KENNETH E. SANTORA • Laboratory of Cellular and Molecular Biology, National Cancer Institute, National Institute of Health, Bethesda, MD ALEXANDER SCHWARZ • Biosphere Medical Inc., Rockland, MA BRYAN JOHN SMITH • Celltech, R&D, Slough, UK VIRGINIA SPENCER • Manitoba Institute of Cell Biology, Manitoba, Canada WAYNE R. SPRINGER • VA San Diego Healthcare System, CA CHRISTOPHER M. STARR • Glyko Inc., Novato, CA KATHRYN L. STONE • Yale Cancer Center Mass Spectrometry Resource and W. M. Keck Foundation Biotechnology Resource Laboratory, New Haven, CT RICHARD A. W. STOTT • Department of Clinical Chemistry, Doncaster Royal Infirmary, South Yorkshire, UK LENNART SVENSSON • Department of Virology, Swedish Institute For Infectious Disease Control, Sweden PATRICIA J. SWEENEY • School of Natural Sciences, Hatfield Polytechnic, University of Hertfordshire, UK DAN S. TAWFIK • Department of Biological Chemistry, the Weizman Institute of Science, Rehovot, Israel JOSEPH THALHAMER • Institute of Chemistry and Biochemistry, Immunology Group, University of Salzburg, Austria JAMES R. THAYER • Dionex Corporation, Life Science Research Group, Sunnyvale, CA GEORGE C. THORNWALL • LECB/NCI-FCRDC, Frederick, MD ROBIN THORPE • Division of Immunobiology, National Institute for Biological Standards and Control, Potters Bar, UK TSUTOMU TSUJI • Hoshi Pharmaceutical College, Tokyo, Japan ROCKY S. TUAN • Department of Orthopaedic Surgery, Thomas Jefferson University Philadelphia, PA
Page 1 and 2: The Protein Protocols Handbook SECO
Page 3 and 4: The Protein Protocols Handbook SECO
Page 5 and 6: Preface The Protein Protocols Handb
Page 7 and 8: viii Contents 14 Identification of
Page 9 and 10: x Contents 47 Conjugation of Fluoro
Page 11 and 12: xii Contents 80 Peptide Mapping by
Page 13 and 14: xiv Contents 112 Sialic Acid Analys
Page 15 and 16: xvi Contents 145 Purification of Ig
Page 17 and 18: Contributors THOMAS E. ADRIAN • D
Page 19: Contributors xxi RUTH R. FRENCH •
Page 23 and 24: UV Absorption 1 PART I QUANTITATION
Page 25 and 26: 4 Aitken and Learmonth 2. Materials
Page 27 and 28: 6 Aitken and Learmonth References 1
Page 29 and 30: 8 Waterborg 3. Method 1. To 0.1 mL
Page 31 and 32: BCA for Protein Quantitation 11 3 T
Page 33 and 34: BCA for Protein Quantitation 13 Tab
Page 35 and 36: The Bradford Method 15 4 The Bradfo
Page 37 and 38: The Bradford Method 17 Fig. 1. Vari
Page 39 and 40: The Bradford Method 19 Table 2 Comp
Page 41 and 42: The Bradford Method 21 13. Kirazov,
Page 43 and 44: 24 Akins and Tuan passes. The side
Page 45 and 46: 26 Akins and Tuan Fig. 2. Effect of
Page 47 and 48: 28 Akins and Tuan protein concentra
Page 49 and 50: 30 Akins and Tuan energy. Too much
Page 51 and 52: 32 Boerner et al. Several approache
Page 53 and 54: 34 Boerner et al. Fig. 2. 3-Nitroty
Page 55 and 56: 36 Boerner et al. containing Tris,
Page 57 and 58: 38 Boerner et al. 2.2. Nitric Acid
Page 59 and 60: 40 Boerner et al. 8. Böhlen, P., S
Page 61 and 62: 42 Aitken and Learmonth 1.2. Measur
Page 63 and 64: 44 Aitken and Learmonth References
Page 65 and 66: 46 Friedrich et al. 2. Materials 2.
Page 67 and 68: 48 Friedrich et al. Fig. 1. Differe
Page 69 and 70: 50 Friedrich et al. References 1. S
Page 71 and 72:
52 Root and Wang Fig. 1. Representa
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54 Root and Wang 4. Kinetic silver
Page 75 and 76:
Gel Electrophoresis of Proteins 57
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Gel Electrophoresis of Proteins 59
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SDS-PAGE 61 11 SDS Polyacrylamide G
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SDS-PAGE 63 their own rates. A more
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SDS-PAGE 65 7. To ensure that the g
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SDS-PAGE 67 close to the dye, and t
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70 Walker 2. Buffers: a. 1.875 M Tr
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72 Walker Fig. 2. Diagrammatic repr
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74 Judd 2. Materials 2.1. Equipment
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76 Judd ber with anode buffer. Remo
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78 Judd 4. Run the gel as described
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SDecS-PAGE of Nucleic Acid Binding
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Page 101 and 102:
Page 103 and 104:
CAT Gel Electrophoresis 87 15 Cetyl
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CAT Gel Electrophoresis 89 Fig. 1.
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CAT Gel Electrophoresis 91 3. CAT s
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CAT Gel Electrophoresis 93 Table 1
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CAT Gel Electrophoresis 95 the tank
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CAT Gel Electrophoresis 97 may be a
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CAT Gel Electrophoresis 99 enzyme p
Page 117 and 118:
CAT Gel Electrophoresis 101 20. Rey
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104 Waterborg Fig. 1. Histones of t
Page 121 and 122:
106 Waterborg 9. Riboflavin-5'-phos
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108 Waterborg 25. Remove the bottom
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110 Waterborg 10. The amount of pro
Page 127 and 128:
AUT Gel for Histones 113 17 Acid-Ur
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AUT Gel for Histones 115 Details of
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AUT Gel for Histones 117 Fig. 2. (A
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AUT Gel for Histones 119 13. Add 0.
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AUT Gel for Histones 121 5. The sep
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AUT Gel for Histones 123 15. Waterb
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126 Walker the cost of preparing IE
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128 Walker 4. Notes 1. The sucrose
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Protein Solubility in 2-D Electroph
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Fractionated Extraction 141 20 Prep
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Fractionated Extraction 143 Fig. 1.
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Fractionated Extraction 145 Table 2
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Fractionated Extraction 147 13. Com
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149 SI + II fraction: liver Brain H
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Fractionated Extraction 151 3. Add
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Fractionated Extraction 153 Fig. 2.
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Fractionated Extraction 155 and bra
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Fractionated Extraction 157 ume are
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160 Bizios 2. Materials 2.1. Equipm
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162 Bizios 5. Sample preparation sh
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164 Gravel Fig. 1. Tube Cell Model
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166 Gravel 3. Fill the lower chambe
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168 Gravel lysis solution A (SDS-DT
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170 Gianazza Fig. 1. Structure of t
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172 Gianazza Table 1 pK Values of I
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174 Gianazza Fig. 2. Course of the
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176 Gianazza Table 3 IPG 4-7 and 6-
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178 Gianazza the gel. Depending on
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180 Gianazza 17. Chiari, M., Pagani
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182 Lopez 3. Slide a grommet onto e
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DIGE 185 25 Difference Gel Electrop
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DIGE 187 2.2. IEF Fig 1. The two cy
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DIGE 189 3. Method 3.1. Sample Solu
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DIGE 191 Strips, the instructions t
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DIGE 193 2. Push the strip down unt
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DIGE 195 4. The presence of Pharmal
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Comparing 2-D Gels on the Internet
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Page 215 and 216:
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Page 219 and 220:
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Page 223 and 224:
Page 225 and 226:
Immunoblotting of 2-DE Separated Pr
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Page 237 and 238:
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Page 241 and 242:
232 Springer Fig. 1. Comparison of
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234 Springer Table 1 Recipe for Var
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Quantification of Proteins on Gels
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Page 249 and 250:
Page 251 and 252:
Rapid Staining with Nile Red 243 30
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Rapid Staining with Nile Red 245 Fi
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Rapid Staining with Nile Red 247 11
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Rapid Staining with Nile Red 249 Re
Page 259 and 260:
252 Fernandez-Patron to years witho
Page 261 and 262:
254 Fernandez-Patron Fig. 2. Revers
Page 263 and 264:
256 Fernandez-Patron spectrometry (
Page 265 and 266:
258 Fernandez-Patron 11. Fernandez-
Page 267 and 268:
260 Choi, Hong, and Yoo Table 1 Lin
Page 269 and 270:
262 Choi, Hong, and Yoo Fig. 2. Mec
Page 271 and 272:
Silver Staining of Proteins 265 33
Page 273 and 274:
Silver Staining of Proteins 267 Fig
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Silver Staining of Proteins 269 3.3
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Silver Staining of Proteins 271 12.
Page 279 and 280:
274 Patton plexes for colorimetric
Page 281 and 282:
276 Patton device (CCD) camera. The
Page 283 and 284:
278 Patton 3.2. Luminescent Detecti
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280 Patton 3.5. Luminescent Detecti
Page 287 and 288:
282 Patton EDTA, pH 9.6 or using SY
Page 289 and 290:
284 Patton filter. Proteins stained
Page 291 and 292:
286 Patton 17. Lim, M., Patton, W.,
Page 293 and 294:
288 Dunn variations in silver stain
Page 295 and 296:
290 Dunn Fig. 1. A 2-DE separation
Page 297 and 298:
292 Dunn 11. Stephens, R. E. (1975)
Page 299 and 300:
Gels Using Eosin Y Stain 295 36 Det
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Gels Using Eosin Y Stain 297 3. An
Page 303 and 304:
300 Jenö and Horst and Coomassie B
Page 305 and 306:
302 Jenö and Horst Fig. 1. Side (A
Page 307 and 308:
304 Jenö and Horst BT1 membrane, o
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Autoradiography and Fluorography 30
Page 311 and 312:
Page 313 and 314:
Page 315 and 316:
Page 317 and 318:
Blotting-Electroblotting 315 PART I
Page 319 and 320:
318 Page and Thorpe 4. Filter paper
Page 322 and 323:
Semidry Protein Blotting 321 40 Pro
Page 324 and 325:
Semidry Protein Blotting 323 2. Mem
Page 326 and 327:
Discontinuous buffer systems Tris-g
Page 328 and 329:
327 Table 2 Membranes Used for Elec
Page 330 and 331:
Semidry Protein Blotting 329 Fig. 2
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Semidry Protein Blotting 331 Table
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Semidry Protein Blotting 333 24 h l
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Protein Blotting 335 41 Protein Blo
Page 338 and 339:
Western Blotting of Basic Proteins
Page 340 and 341:
Page 342 and 343:
Page 344 and 345:
344 Wisdom 4. Glutaraldehyde. 5. 50
Page 346 and 347:
346 Wisdom 3. Methods 1. Dissolve 1
Page 348 and 349:
348 Wisdom 3. Dialyze the modified
Page 350 and 351:
350 Wisdom 6. Store the labeled ant
Page 352 and 353:
352 Mao terminus. The ε-amino grou
Page 354 and 355:
354 Mao conjugate with optimal modi
Page 356 and 357:
356 Haugland and You Fig. 1. Struct
Page 358 and 359:
358 Haugland and You Because of its
Page 360 and 361:
360 Haugland and You 5. Dissolve 10
Page 362 and 363:
362 Haugland and You ing with one a
Page 364 and 365:
Preparation of Avidin Conjugates 36
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Page 368 and 369:
Page 370 and 371:
Page 372 and 373:
Page 374 and 375:
Staining with MDPF 375 50 MDPF Stai
Page 376 and 377:
Staining with MDPF 377 transillumin
Page 378 and 379:
Staining with MDPF 379 3. Alba, F.
Page 380 and 381:
382 Root and Wang suspension become
Page 382 and 383:
384 Root and Wang Fig. 1. Schematic
Page 384 and 385:
Blots Using Direct Blue 71 387 52 D
Page 386 and 387:
Blots Using Direct Blue 71 389 3.2.
Page 388 and 389:
Blots Using Direct Blue 71 391 Fig.
Page 390 and 391:
Immunogold 393 53 Protein Staining
Page 392 and 393:
Immunogold 395 Fig. 2. Schematic re
Page 394 and 395:
Immunogold 397 6. AuroProbe BLplus
Page 396 and 397:
Immunogold 399 Fig. 5. Comparison o
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Immunogold 401 Table 2 Effect of Te
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Immunogold 403 15. AuroDye forte is
Page 402 and 403:
Immunoblotting Using Secondary Liga
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Page 406 and 407:
Page 408 and 409:
Page 410 and 411:
Page 412 and 413:
Avidin-or Streptavidin-Biotin 415 5
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Avidin-or Streptavidin-Biotin 417 2
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Avidin-or Streptavidin-Biotin 419 R
Page 418 and 419:
422 Copse and Fowler substrate to a
Page 420 and 421:
424 Copse and Fowler 2. Orbital sha
Page 422 and 423:
426 Copse and Fowler 4. Notes 4.1.
Page 424 and 425:
428 Copse and Fowler 18. DDAO-phosp
Page 426 and 427:
430 Dickinson and Fowler the advant
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432 Dickinson and Fowler 2. Membran
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434 Dickinson and Fowler Fig. 2. (A
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436 Dickinson and Fowler biotinylat
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Reutilization of Western Blots 439
Page 437 and 438:
Page 439 and 440:
Page 441 and 442:
Page 443 and 444:
Page 445 and 446:
Page 447 and 448:
Page 449 and 450:
Carboxymethylation of Cysteine 453
Page 451 and 452:
456 Aitken and Learmonth 11. Microd
Page 453 and 454:
458 Aitken and Learmonth Table 1 El
Page 455 and 456:
460 Aitken and Learmonth 4. Notes 1
Page 457 and 458:
462 Ward Fig. 1. Amino acid sequenc
Page 459 and 460:
Chemical Modifications of Proteins
Page 461 and 462:
Chemical Modifications of Proteins
Page 463 and 464:
470 Tawfik 3. Method 3.1. Nitration
Page 465 and 466:
472 Tawfik preparation by pH-depend
Page 467 and 468:
474 Tawfik 4. Notes 1. The concentr
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476 Tawfik 2. A molar excess of p-h
Page 471 and 472:
478 Tawfik 3. Method 1. Add the gly
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480 Tawfik 3. Method 1. Dissolve th
Page 475 and 476:
482 Tawfik 4. Notes 1. At neutral a
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484 Tawfik 4. Notes 1. Release of t
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486 Smith 6. Equipment includes a n
Page 481 and 482:
488 Smith 3. Although the specifici
Page 483 and 484:
490 Smith those bearing a prolyl re
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Cleavage at Tryptophanyl-x Bonds 49
Page 487 and 488:
Alternative conditions for rapid re
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Cleavage at Tryptophanyl-x Bonds 49
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Cleavage at Aspartyl-X Bonds 499 73
Page 493 and 494:
Cleavage at Aspartyl-X Bonds 501 pr
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Cleavage at Cysteinyl-x Bonds 503 7
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Cleavage at Cysteinyl-x Bonds 505 F
Page 499 and 500:
Cleavage at Asparaginyl-Glycyl Bond
Page 501 and 502:
Cleavage at Asparaginyl-Glycyl Bond
Page 503 and 504:
Enzymatic Digestion 511 76 Enzymati
Page 505 and 506:
Enzymatic Digestion 513 then remove
Page 507 and 508:
Enzymatic Digestion 515 Fig. 1. In-
Page 509 and 510:
Enzymatic Digestion 517 In the case
Page 511 and 512:
Enzymatic Digestion 519 Another app
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Enzymatic Digestion 521 11. Modific
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524 Table 1 Digestion Buffers Recip
Page 517 and 518:
526 Fernandez and Mische Fig. 1. Pe
Page 519 and 520:
528 Fernandez and Mische 3. Excise
Page 521 and 522:
530 Fernandez and Mische 3. The lar
Page 523 and 524:
532 Fernandez and Mische membrane a
Page 525 and 526:
534 Stone and Williams 2. Materials
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536 Stone and Williams In those few
Page 529 and 530:
538 Stone and Williams (while maint
Page 531 and 532:
540 Stone and Williams Biochemistry
Page 533 and 534:
2-D TLE-TLC Mapping 543 79 Peptide
Page 535 and 536:
2-D TLE-TLC Mapping 545 12. 0.25% N
Page 537 and 538:
2-D TLE-TLC Mapping 547 3. Incubate
Page 539 and 540:
2-D TLE-TLC Mapping 549 Table 1 Cle
Page 541 and 542:
2-D TLE-TLC Mapping 551 Fig. 1. Exa
Page 543 and 544:
SDS-PAGE Mapping 553 80 Peptide Map
Page 545 and 546:
SDS-PAGE Mapping 555 3. Overlay sam
Page 547 and 548:
SDS-PAGE Mapping 557 Fig. 1. Exampl
Page 549 and 550:
560 Judd Fig. 1. Example of peptide
Page 551 and 552:
Protein Hydrolysates 563 82 Product
Page 553 and 554:
Protein Hydrolysates 565 2. Pronase
Page 555 and 556:
Amino Acid Analysis with Marfey’s
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Page 559 and 560:
Page 561 and 562:
HPSEC 573 84 Molecular Weight Estim
Page 563 and 564:
HPSEC 575 Table 1 Protein Standards
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HPSEC 577 Fig. 2. Plot of K d vs lo
Page 567 and 568:
HPSEC 579 with care (see suppliers
Page 569 and 570:
582 Aitken larly good resolution de
Page 571 and 572:
Disulfide-Linked Peptide Detection
Page 573 and 574:
Disulfide-Linked Peptide Detection
Page 575 and 576:
Disulfide Bridges 589 87 Diagonal E
Page 577 and 578:
Disulfide Bridges 591 3. Spot the s
Page 579 and 580:
Disulfide Bridges 593 2. The moveme
Page 581 and 582:
596 Aitken and Learmonth 6. Finally
Page 583 and 584:
598 Aitken and Learmonth Fig. 1. (A
Page 585 and 586:
600 Aitken and Learmonth 2.4. Elect
Page 587 and 588:
602 Aitken and Learmonth 6. Takahas
Page 589 and 590:
604 Colyer of the protein of intere
Page 591 and 592:
606 Colyer 8. After 16 h of exposur
Page 593 and 594:
608 Colyer stoichiometry, since it
Page 595 and 596:
610 Bonenfant, Mini, and Jenö indi
Page 597 and 598:
612 Bonenfant, Mini, and Jenö for
Page 599 and 600:
614 Bonenfant, Mini, and Jenö incl
Page 601 and 602:
616 Bonenfant, Mini, and Jenö Fig.
Page 603 and 604:
618 Bonenfant, Mini, and Jenö Fig.
Page 605 and 606:
620 Bonenfant, Mini, and Jenö up t
Page 607 and 608:
622 Bonenfant, Mini, and Jenö 5. L
Page 609 and 610:
624 Weber and McFadden Fig. 1. Sche
Page 611 and 612:
626 Weber and McFadden 4. Using a p
Page 613 and 614:
628 Weber and McFadden Fig. 2. Comp
Page 615 and 616:
630 Weber and McFadden Fig. 4. Coom
Page 617 and 618:
Protein Palmitoylation 633 93 Analy
Page 619 and 620:
Protein Palmitoylation 635 Fig. 1.
Page 621 and 622:
Protein Palmitoylation 637 1. Fix p
Page 623 and 624:
Protein Palmitoylation 639 7. Mumby
Page 625 and 626:
Fig. 1. Structure of the natural an
Page 627 and 628:
644 Corsini 8. Simvastatin in its l
Page 629 and 630:
646 Corsini Fig. 3. Schematic for t
Page 631 and 632:
648 Corsini Table 1 Mevalonate, Pre
Page 633 and 634:
650 Corsini Fig. 5. Metabolic label
Page 635 and 636:
652 Corsini Fig. 7. Two-dimensional
Page 637 and 638:
654 Corsini 7. When prenylated prot
Page 639 and 640:
656 Corsini 24. Danesi, R., McLella
Page 641 and 642:
658 Andres et al. Fig. 1. Proposed
Page 643 and 644:
660 Andres et al. Fig. 2. SDS-PAGE
Page 645 and 646:
662 Andres et al. Fig. 4. Chromatog
Page 647 and 648:
664 Andres et al. Fig. 6. Specific
Page 649 and 650:
666 Andres et al. 10. Residual orga
Page 651 and 652:
668 Andres et al. 2. The metabolica
Page 653 and 654:
670 Andres et al. 3. Clarke, S. (19
Page 655 and 656:
Phosphopeptide Mapping 673 96 2-D P
Page 657 and 658:
Phosphopeptide Mapping 675 3. Add 1
Page 659 and 660:
Phosphopeptide Mapping 677 Fig. 1.
Page 661 and 662:
Phosphopeptide Mapping 679 until th
Page 663 and 664:
Protein Mutations by MS 681 97 Dete
Page 665 and 666:
Protein Mutations by MS 683 protein
Page 667 and 668:
Protein Mutations by MS 685 Fig. 2.
Page 669 and 670:
Page 671 and 672:
Page 673 and 674:
Protein Mutations by MS 691 Fig. 10
Page 675 and 676:
Nanoelectrospray MS/MS for Peptide
Page 677 and 678:
Page 679 and 680:
Page 681 and 682:
Page 683 and 684:
Page 685 and 686:
Page 687 and 688:
Page 689 and 690:
Page 691 and 692:
Page 693 and 694:
MALDI-MS for Protein Identification
Page 695 and 696:
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Page 701 and 702:
Page 703 and 704:
Page 705 and 706:
Page 707 and 708:
Page 709 and 710:
Page 711 and 712:
Page 713 and 714:
Page 715 and 716:
Protein Ladder Sequencing 733 100 P
Page 717 and 718:
Protein Ladder Sequencing 735 Prote
Page 719 and 720:
Protein Ladder Sequencing 737 3. Ex
Page 721 and 722:
Protein Ladder Sequencing 739 2. Wa
Page 723 and 724:
742 Hennig sequences (see Subheadin
Page 725 and 726:
744 Hennig In the age of genomics,
Page 727 and 728:
746 Hennig last defined (last in th
Page 729 and 730:
748 Spencer and Davie Fig. 1. Two-d
Page 731 and 732:
750 Spencer and Davie 4. Notes 1. T
Page 733 and 734:
Proteins Cross-linked to DNA by For
Page 735 and 736:
Page 737 and 738:
Page 739 and 740:
Glycoprotein Detection 761 104 Dete
Page 741 and 742:
Glycoprotein Detection 763 Schiff
Page 743 and 744:
Glycoprotein Detection 765 3.1.1. G
Page 745 and 746:
Glycoprotein Detection 767 Table 1
Page 747 and 748:
Glycoprotein Detection 769 5. Resus
Page 749 and 750:
Glycoprotein Detection 771 Fig. 1.
Page 751 and 752:
Detection of Glycosylated Proteins
Page 753 and 754:
Page 755 and 756:
Page 757 and 758:
780 Gravel Fig. 1. Glycoprotein blo
Page 759 and 760:
782 Gravel Fig. 3. Glycoprotein blo
Page 761 and 762:
784 Table 1 Glycans N-Glycosidicall
Page 763 and 764:
786 Table 3 Specificity of Lectins
Page 765 and 766:
788 Table 3 Specificity of Lectins
Page 767 and 768:
790 Gravel dimethylformamide. These
Page 769 and 770:
792 Gravel 3. Montreuil, J., Bouque
Page 771 and 772:
794 Gravel
Page 773 and 774:
796 Goodarzi, Fotinopoulou, Turner
Page 775 and 776:
Page 777 and 778:
Page 779 and 780:
Page 781 and 782:
804 Hounsell, Davies, and Smith 2.
Page 783 and 784:
Page 785 and 786:
Monosaccharide Analysis by GC 809 1
Page 787 and 788:
Linkage and Substitution Patterns 8
Page 789 and 790:
Linkage and Substitution Patterns 8
Page 791 and 792:
Page 793 and 794:
Page 795 and 796:
820 Hounsell, Davis, and Smith 6. T
Page 797 and 798:
Page 799 and 800:
824 Mizuochi and Hounsell 3. Method
Page 801 and 802:
826 Mizuochi and Hounsell Reference
Page 803 and 804:
Page 805 and 806:
Page 807 and 808:
Page 809 and 810:
834 Weitzhandler et al. alditols (1
Page 811 and 812:
836 Weitzhandler et al. Fig. 1. HPA
Page 813 and 814:
838 Weitzhandler et al. 2. Add 0.1
Page 815 and 816:
Microassay of Protein Glycosylation
Page 817 and 818:
Page 819 and 820:
Page 821 and 822:
Page 823 and 824:
Page 825 and 826:
Carbohydrate Electrophoresis 851 12
Page 827 and 828:
Carbohydrate Electrophoresis 853 2.
Page 829 and 830:
Carbohydrate Electrophoresis 855 Pl
Page 831 and 832:
Page 833 and 834:
Carbohydrate Electrophoresis 859 Fi
Page 835 and 836:
Carbohydrate Electrophoresis 861 wa
Page 837 and 838:
Page 839 and 840:
866 Merry blly less expensive than
Page 841 and 842:
868 Merry 8. Whatman no. 3 chromato
Page 843 and 844:
870 Merry 4. Dialyze for a minimum
Page 845 and 846:
872 Merry 9. Leave for 5 min and un
Page 847 and 848:
874 Merry 2. Column: Reverse-phase
Page 849 and 850:
Table 1 Incubation Conditions for E
Page 851 and 852:
878 Merry 3.7. Exoglycosidase Diges
Page 853 and 854:
880 Merry Fig. 6. Example of exogly
Page 855 and 856:
882 Merry 17. Rice, K. G., Takahash
Page 857 and 858:
Glycoprofiling and SPR 885 124 Glyc
Page 859 and 860:
Glycoprofiling and SPR 887 2. Mater
Page 861 and 862:
Glycoprofiling and SPR 889 Fig. 3.
Page 863 and 864:
Glycoprofiling and SPR 891 α2-3 Ne
Page 865 and 866:
894 Turnbull Fig. 1. Basic principl
Page 867 and 868:
896 Turnbull 13. Enzyme buffer (0.2
Page 869 and 870:
898 Turnbull Table 1 Exoenzymes for
Page 871 and 872:
900 Turnbull Fig. 3. IGS of a hepar
Page 873 and 874:
902 Turnbull 4. Notes 1. Using larg
Page 875 and 876:
904 Turnbull 20. Rice, K., Rottink,
Page 877 and 878:
906 Hooker and James 3. High-perfor
Page 879 and 880:
908 Hooker and James Fig. 1. Whole
Page 881 and 882:
910 Hooker and James at individual
Page 883 and 884:
912 Hooker and James Fig. 4. Sialyl
Page 885 and 886:
914 Hooker and James 16. Ashton, D.
Page 887 and 888:
Lectin Affinity Chromatography 917
Page 889 and 890:
Page 891 and 892:
Page 893 and 894:
Page 895 and 896:
Page 897 and 898:
Page 899 and 900:
Page 901 and 902:
Page 903 and 904:
Antibody Production 935 128 Antibod
Page 905 and 906:
Antibody Production 937 1.1. Donor
Page 907 and 908:
Antibody Production 939 2. Material
Page 909 and 910:
Production of Anitbodies Using Prot
Page 911 and 912:
Production of Anitbodies Using Prot
Page 913 and 914:
Production of Polyclonal Antibodies
Page 915 and 916:
Page 917 and 918:
Page 919 and 920:
Page 921 and 922:
Peptide Conjugation and Immunizatio
Page 923 and 924:
Page 925 and 926:
Page 927 and 928:
Page 929 and 930:
Page 931 and 932:
964 Bailey is oxidized by chloramin
Page 933 and 934:
The Lactoperoxidase Method 967 133
Page 935 and 936:
The Bolton and Hunter Method 969 13
Page 937 and 938:
Radioiodination Using IODO-GEN 971
Page 939 and 940:
Page 941 and 942:
Page 943 and 944:
Page 945 and 946:
980 Bailey 3. Specific antiseum to
Page 947 and 948:
982 Bailey Amount of radioactivity
Page 949 and 950:
984 Page and Thorpe 2. Centrifuge s
Page 951 and 952:
DEAE-Sepharose Chromatography 987 1
Page 953 and 954:
IgG Purification with Ion-Exchange
Page 955 and 956:
PEG Precipitation 991 141 Purificat
Page 957 and 958:
994 Page and Thorpe 2. Materials 1.
Page 959 and 960:
996 Dolman and Thorpe Fig. 1. Typic
Page 961 and 962:
Antigen-Ligand Columns 999 144 Puri
Page 963 and 964:
Antigen-Ligand Columns 1001 4. When
Page 965 and 966:
1004 Page and Thorpe 7. Filter and
Page 967 and 968:
1006 Page and Thorpe Fig. 1. SDS-PA
Page 969 and 970:
Purification of IgY from Chicken Eg
Page 971 and 972:
Purification of IgY from Chicken Eg
Page 973 and 974:
1014 Fassina et al. limit the use o
Page 975 and 976:
1016 Fassina et al. 2. Materials Ta
Page 977 and 978:
1018 Fassina et al. 10. Filter the
Page 979 and 980:
1020 Fassina et al. 1. Dilute sampl
Page 981 and 982:
1022 Fassina et al. analysis indica
Page 983 and 984:
1024 Fassina et al. 6. Khayam-Bashi
Page 985 and 986:
1026 Hammerl et al. down, with the
Page 987 and 988:
1028 Hammerl et al. 1. Clean glass
Page 989 and 990:
1030 Hammerl et al. Fig. 1. Experim
Page 991 and 992:
1032 Hammerl et al. can “corrode
Page 993 and 994:
Single-Chain Antibodies 1035 150 Ba
Page 995 and 996:
Single-Chain Antibodies 1037 ing th
Page 997 and 998:
Single-Chain Antibodies 1039 6. Mag
Page 999 and 1000:
Single-Chain Antibodies 1041 XL1-Bl
Page 1001 and 1002:
Single-Chain Antibodies 1043 fore,
Page 1003 and 1004:
Single-Chain Antibodies 1045 16. Ho
Page 1005 and 1006:
Enzymatic Digestion of MAbs 1047 15
Page 1007 and 1008:
Enzymatic Digestion of MAbs 1049 2.
Page 1009 and 1010:
Enzymatic Digestion of MAbs 1051 2.
Page 1011 and 1012:
Making Bispecific Antibodies 1053 1
Page 1013 and 1014:
Page 1015 and 1016:
Making Bispecific Antibodies 1057 F
Page 1017 and 1018:
Phage Display 1059 153 Phage Displa
Page 1019 and 1020:
Phage Display 1061 5000, 1 mL of 2-
Page 1021 and 1022:
Phage Display 1063 11. Agarose top:
Page 1023 and 1024:
Phage Display 1065 3.1.3.2. AFFINIT
Page 1025 and 1026:
Phage Display 1067 8. Shake vigorou
Page 1027 and 1028:
Phage Display 1069 4. Notes 1. Fila
Page 1029 and 1030:
Phage Display 1071 9. Sengupta J.,
Page 1031 and 1032:
Selection by Panning 1073 154 Scree
Page 1033 and 1034:
Selection by Panning 1075 Fig. 1. F
Page 1035 and 1036:
Selection by Panning 1077 12. Centr
Page 1037 and 1038:
Selection by Panning 1079 4. Notes
Page 1039 and 1040:
Selection by Panning 1081 33. The
Page 1041 and 1042:
Antigen Measurement Using ELISA 108
Page 1043 and 1044:
Page 1045 and 1046:
Page 1047 and 1048:
Enhanced Chemiluminescence 1089 156
Page 1049 and 1050:
Enhanced Chemiluminescence 1091 Tab
Page 1051 and 1052:
Enhanced Chemiluminescence 1093 is
Page 1053 and 1054:
Enhanced Chemiluminescence 1095 lig
Page 1055 and 1056:
Immunoprecipitation 1097 157 Immuno
Page 1057 and 1058:
Immunoprecipitation 1099 Table 2 Pr
Page 1059 and 1060:
Immunoprecipitation 1101 oligosacch
Page 1061 and 1062:
Immunoprecipitation 1103 6. Very ge
Page 1063 and 1064:
Immunoprecipitation 1105 2. Dry the
Page 1065 and 1066:
Short Chapter Title 1107 PART VIII
Page 1067 and 1068:
1110 Page and Thorpe final quantity
Page 1069 and 1070:
1112 Page and Thorpe 2. Materials 1
Page 1071 and 1072:
Page 1073 and 1074:
161 From: The Protein Protocols Han
Page 1075 and 1076:
162 From: The Protein Protocols Han
Page 1077 and 1078:
Affinity Purification of Monoclonal
Page 1079 and 1080:
Page 1081 and 1082:
Page 1083 and 1084:
Page 1085 and 1086:
An Efficient Method for MAb Product
Page 1087 and 1088:
Page 1089 and 1090:
Page 1091 and 1092:
Page 1093 and 1094:
Page 1095 and 1096:
Index 1139 Index A Absorbance (UV),
Page 1097 and 1098:
Index 1141 Electrophoresis of antib
Page 1099 and 1100:
Index 1143 of glycoproteins, 905-91
Page 1101 and 1102:
Index 1145 India ink, 338 lectin st
Page 1103:
The Protein Protocols Handbook Seco
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