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Calconcarboxylic Acid 263 2. Merril, C. R., Goldman, D., Sedman, S., and Ebert, M. (1980) Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science 211, 1437,1438. 3. Schetters, H. and McLeod, B. (1979) Simultaneous isolation of major viral proteins in one step. Analyt. Biochem. 98, 329–334. 4. Jakowski, G. and Liew, C. C. (1980) Fluorescamine staining of nonhistone chromatin proteins as revealed by two-dimensional polyacrylamide gel electrophoresis. Analyt. Biochem. 102, 321–325. 5. Weiderkamm, E., Wallach, D. F. H., and Fluckiger, R. (1973) A new sensitive, rapid fluorescence technique for the determination of proteins in gel electrophoresis and in solution. Analyt. Biochem. 54, 102–114. 6. Gabriel, O. (1971) Locating enzymes on gels in Methods in Enzymology, vol. 22 (Colowick, S. P. and Kaplan, N. O., eds.), Academic, New York, pp. 363–367. 7. O’Farrell, P. H. (1975) High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250, 4007–4021. 8. Diezel, W., Kopperschlager, G., and Hofmann, E. (1972) An improved procedure for protein staining in polyacrylamide gels with a new type of Coomassie Brilliant Blue. Analyt. Biochem. 48, 617–620. 9. Zehr, B. D., Savin, T. J., and Hall, R. E. (1989) A one-step low background Coomassie staining procedure for polyacrylamide gels. Analyt. Biochem. 182, 157–159. 10. Hames, B. D. and Rickwood, D. (1990) Analysis of gels following electrophoresis, in Gel Electrophoresis of Proteins: A Practical Approach, IRL, Oxford, UK, pp. 52–81. 11. Switzer, R. C., Merril, C. R., and Shifrin, S. (1979) A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels. Analyt. Biochem. 98, 231–237. 12. Ohsawa, K. and Ebata, N. (1983) Silver stain for detecting 10-femtogram quantities of protein after polyacrylamide gel electrophoresis. Analyt. Biochem. 135, 409–415. 13. Hong, H. Y., Yoo, G. S., and Choi, J. K. (1993) Detection of proteins on polyacrylamide gels using calconcarboxylic acid. Analyt. Biochem. 214, 96–99. 14. Borejdo, J. and Flynn, C. (1984) Electrophoresis in the presence of Coomassie Brilliant Blue R-250 stains polyacrylamide gels during protein fractionation. Analyt. Biochem. 140, 84–86.
Silver Staining of Proteins 265 33 Detection of Proteins in Polyacrylamide Gels by Silver Staining Michael J. Dunn 1. Introduction The versatility and resolving capacity of polyacrylamide gel electrophoresis has resulted in this group of methods becoming the most popular for the analysis of patterns of gene expression in a wide variety of complex systems. These techniques are often used to characterize protein purity and to monitor the various steps in a protein purification process. Moreover, two-dimensional polyacrylamide gel electrophoresis (2-DE) remains the core technology of choice for separating complex protein mixtures in the majority of proteome projects (1, and see Chapter 22). This is due to its unrivalled power to separate simultaneously thousands of proteins and the relative ease with which proteins from 2-D gels can be identified and characterized using highly sensitive microchemical methods (2), particularly those based on mass spectrometry (3). Gel electrophoresis is now one of the most important methods of protein purification for subsequent identification and characterization. Coomassie Brilliant Blue R-250 (CBB R-250) has been used for many years as a general protein stain following gel electrophoresis. However, the trend toward the use of thinner gels and the need to detect small amount of protein within single bands or spots resolved by one- or two-dimensional electrophoresis have necessitated the development of more sensitive detection methods (4). The ability of silver to develop images was discovered in the mid-17th century and this property was exploited in the development of photography, followed by its use in histological procedures. Silver staining for the detection of proteins following gel electrophoresis was first reported in 1979 by Switzer et al. (5), resulting in a major increase in the sensitivity of protein detection. More than 100 publications have subsequently appeared describing variations in silver staining methodology (4,6). This group of procedures is generally accepted to be around 100 times more sensitive than methods using CBB R-250, being able to detect low nanogram amounts of protein per gel band or spot. All silver staining procedures depend on the reduction of ionic silver to its metallic form, but the precise mechanism involved in the staining of proteins has not been fully established. It has been proposed that silver cations complex with protein amino groups, From: The Protein Protocols Handbook, 2nd Edition Edited by: J. M. Walker © Humana Press Inc., Totowa, NJ 265
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The Protein Protocols Handbook SECO
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The Protein Protocols Handbook SECO
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Preface The Protein Protocols Handb
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viii Contents 14 Identification of
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x Contents 47 Conjugation of Fluoro
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xii Contents 80 Peptide Mapping by
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xiv Contents 112 Sialic Acid Analys
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xvi Contents 145 Purification of Ig
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Contributors THOMAS E. ADRIAN • D
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Contributors xxi RUTH R. FRENCH •
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Contributors xxiii STEFANIE A. NELS
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UV Absorption 1 PART I QUANTITATION
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4 Aitken and Learmonth 2. Materials
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6 Aitken and Learmonth References 1
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8 Waterborg 3. Method 1. To 0.1 mL
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BCA for Protein Quantitation 11 3 T
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BCA for Protein Quantitation 13 Tab
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The Bradford Method 15 4 The Bradfo
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The Bradford Method 17 Fig. 1. Vari
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The Bradford Method 19 Table 2 Comp
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The Bradford Method 21 13. Kirazov,
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24 Akins and Tuan passes. The side
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26 Akins and Tuan Fig. 2. Effect of
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28 Akins and Tuan protein concentra
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30 Akins and Tuan energy. Too much
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32 Boerner et al. Several approache
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34 Boerner et al. Fig. 2. 3-Nitroty
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36 Boerner et al. containing Tris,
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38 Boerner et al. 2.2. Nitric Acid
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40 Boerner et al. 8. Böhlen, P., S
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42 Aitken and Learmonth 1.2. Measur
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44 Aitken and Learmonth References
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46 Friedrich et al. 2. Materials 2.
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48 Friedrich et al. Fig. 1. Differe
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50 Friedrich et al. References 1. S
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52 Root and Wang Fig. 1. Representa
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54 Root and Wang 4. Kinetic silver
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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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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
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CAT Gel Electrophoresis 101 20. Rey
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104 Waterborg Fig. 1. Histones of t
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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
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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 219 and 220: Comparing 2-D Gels on the Internet
Page 225 and 226: Immunoblotting of 2-DE Separated Pr
Page 241 and 242: 232 Springer Fig. 1. Comparison of
Page 243 and 244: 234 Springer Table 1 Recipe for Var
Page 245 and 246: Quantification of Proteins on Gels
Page 251 and 252: Rapid Staining with Nile Red 243 30
Page 253 and 254: Rapid Staining with Nile Red 245 Fi
Page 255 and 256: Rapid Staining with Nile Red 247 11
Page 257 and 258: 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: 262 Choi, Hong, and Yoo Fig. 2. Mec
Page 273 and 274: Silver Staining of Proteins 267 Fig
Page 275 and 276: Silver Staining of Proteins 269 3.3
Page 277 and 278: 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
Page 285 and 286: 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
Page 301 and 302: 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
Page 309 and 310: Autoradiography and Fluorography 30
Page 317 and 318: Blotting-Electroblotting 315 PART I
Page 319 and 320: 318 Page and Thorpe 4. Filter paper
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Semidry Protein Blotting 321 40 Pro
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Semidry Protein Blotting 323 2. Mem
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Discontinuous buffer systems Tris-g
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327 Table 2 Membranes Used for Elec
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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
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Western Blotting of Basic Proteins
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344 Wisdom 4. Glutaraldehyde. 5. 50
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346 Wisdom 3. Methods 1. Dissolve 1
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348 Wisdom 3. Dialyze the modified
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350 Wisdom 6. Store the labeled ant
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352 Mao terminus. The ε-amino grou
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354 Mao conjugate with optimal modi
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356 Haugland and You Fig. 1. Struct
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358 Haugland and You Because of its
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360 Haugland and You 5. Dissolve 10
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362 Haugland and You ing with one a
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Preparation of Avidin Conjugates 36
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Staining with MDPF 375 50 MDPF Stai
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Staining with MDPF 377 transillumin
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Staining with MDPF 379 3. Alba, F.
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382 Root and Wang suspension become
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384 Root and Wang Fig. 1. Schematic
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Blots Using Direct Blue 71 387 52 D
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Blots Using Direct Blue 71 389 3.2.
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Blots Using Direct Blue 71 391 Fig.
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Immunogold 393 53 Protein Staining
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Immunogold 395 Fig. 2. Schematic re
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Immunogold 397 6. AuroProbe BLplus
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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
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Immunoblotting Using Secondary Liga
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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
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422 Copse and Fowler substrate to a
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424 Copse and Fowler 2. Orbital sha
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426 Copse and Fowler 4. Notes 4.1.
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428 Copse and Fowler 18. DDAO-phosp
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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
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Carboxymethylation of Cysteine 453
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456 Aitken and Learmonth 11. Microd
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458 Aitken and Learmonth Table 1 El
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460 Aitken and Learmonth 4. Notes 1
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462 Ward Fig. 1. Amino acid sequenc
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Chemical Modifications of Proteins
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Chemical Modifications of Proteins
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470 Tawfik 3. Method 3.1. Nitration
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472 Tawfik preparation by pH-depend
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474 Tawfik 4. Notes 1. The concentr
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476 Tawfik 2. A molar excess of p-h
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478 Tawfik 3. Method 1. Add the gly
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480 Tawfik 3. Method 1. Dissolve th
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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
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488 Smith 3. Although the specifici
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490 Smith those bearing a prolyl re
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Cleavage at Tryptophanyl-x Bonds 49
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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
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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
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Cleavage at Asparaginyl-Glycyl Bond
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Cleavage at Asparaginyl-Glycyl Bond
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Enzymatic Digestion 511 76 Enzymati
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Enzymatic Digestion 513 then remove
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Enzymatic Digestion 515 Fig. 1. In-
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Enzymatic Digestion 517 In the case
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Enzymatic Digestion 519 Another app
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Enzymatic Digestion 521 11. Modific
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524 Table 1 Digestion Buffers Recip
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526 Fernandez and Mische Fig. 1. Pe
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528 Fernandez and Mische 3. Excise
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530 Fernandez and Mische 3. The lar
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532 Fernandez and Mische membrane a
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534 Stone and Williams 2. Materials
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536 Stone and Williams In those few
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538 Stone and Williams (while maint
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540 Stone and Williams Biochemistry
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2-D TLE-TLC Mapping 543 79 Peptide
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2-D TLE-TLC Mapping 545 12. 0.25% N
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2-D TLE-TLC Mapping 547 3. Incubate
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2-D TLE-TLC Mapping 549 Table 1 Cle
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2-D TLE-TLC Mapping 551 Fig. 1. Exa
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SDS-PAGE Mapping 553 80 Peptide Map
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SDS-PAGE Mapping 555 3. Overlay sam
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SDS-PAGE Mapping 557 Fig. 1. Exampl
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560 Judd Fig. 1. Example of peptide
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Protein Hydrolysates 563 82 Product
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Protein Hydrolysates 565 2. Pronase
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Amino Acid Analysis with Marfey’s
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HPSEC 573 84 Molecular Weight Estim
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HPSEC 575 Table 1 Protein Standards
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HPSEC 577 Fig. 2. Plot of K d vs lo
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HPSEC 579 with care (see suppliers
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582 Aitken larly good resolution de
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Disulfide-Linked Peptide Detection
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Disulfide-Linked Peptide Detection
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Disulfide Bridges 589 87 Diagonal E
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Disulfide Bridges 591 3. Spot the s
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Disulfide Bridges 593 2. The moveme
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596 Aitken and Learmonth 6. Finally
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598 Aitken and Learmonth Fig. 1. (A
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600 Aitken and Learmonth 2.4. Elect
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602 Aitken and Learmonth 6. Takahas
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604 Colyer of the protein of intere
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606 Colyer 8. After 16 h of exposur
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608 Colyer stoichiometry, since it
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610 Bonenfant, Mini, and Jenö indi
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612 Bonenfant, Mini, and Jenö for
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614 Bonenfant, Mini, and Jenö incl
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616 Bonenfant, Mini, and Jenö Fig.
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618 Bonenfant, Mini, and Jenö Fig.
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620 Bonenfant, Mini, and Jenö up t
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622 Bonenfant, Mini, and Jenö 5. L
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624 Weber and McFadden Fig. 1. Sche
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626 Weber and McFadden 4. Using a p
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628 Weber and McFadden Fig. 2. Comp
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630 Weber and McFadden Fig. 4. Coom
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Protein Palmitoylation 633 93 Analy
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Protein Palmitoylation 635 Fig. 1.
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Protein Palmitoylation 637 1. Fix p
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Protein Palmitoylation 639 7. Mumby
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Fig. 1. Structure of the natural an
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644 Corsini 8. Simvastatin in its l
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646 Corsini Fig. 3. Schematic for t
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648 Corsini Table 1 Mevalonate, Pre
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650 Corsini Fig. 5. Metabolic label
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652 Corsini Fig. 7. Two-dimensional
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654 Corsini 7. When prenylated prot
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656 Corsini 24. Danesi, R., McLella
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658 Andres et al. Fig. 1. Proposed
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660 Andres et al. Fig. 2. SDS-PAGE
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662 Andres et al. Fig. 4. Chromatog
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664 Andres et al. Fig. 6. Specific
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666 Andres et al. 10. Residual orga
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668 Andres et al. 2. The metabolica
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670 Andres et al. 3. Clarke, S. (19
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Phosphopeptide Mapping 673 96 2-D P
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Phosphopeptide Mapping 675 3. Add 1
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Phosphopeptide Mapping 677 Fig. 1.
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Phosphopeptide Mapping 679 until th
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Protein Mutations by MS 681 97 Dete
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Protein Mutations by MS 683 protein
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Protein Mutations by MS 685 Fig. 2.
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Protein Mutations by MS 691 Fig. 10
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Nanoelectrospray MS/MS for Peptide
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MALDI-MS for Protein Identification
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Page 713 and 714:
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Protein Ladder Sequencing 733 100 P
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Protein Ladder Sequencing 735 Prote
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Protein Ladder Sequencing 737 3. Ex
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Protein Ladder Sequencing 739 2. Wa
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742 Hennig sequences (see Subheadin
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744 Hennig In the age of genomics,
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746 Hennig last defined (last in th
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748 Spencer and Davie Fig. 1. Two-d
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750 Spencer and Davie 4. Notes 1. T
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Proteins Cross-linked to DNA by For
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Page 737 and 738:
Page 739 and 740:
Glycoprotein Detection 761 104 Dete
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Glycoprotein Detection 763 Schiff
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Glycoprotein Detection 765 3.1.1. G
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Glycoprotein Detection 767 Table 1
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Glycoprotein Detection 769 5. Resus
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Glycoprotein Detection 771 Fig. 1.
Page 751 and 752:
Detection of Glycosylated Proteins
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Page 757 and 758:
780 Gravel Fig. 1. Glycoprotein blo
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782 Gravel Fig. 3. Glycoprotein blo
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784 Table 1 Glycans N-Glycosidicall
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786 Table 3 Specificity of Lectins
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788 Table 3 Specificity of Lectins
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790 Gravel dimethylformamide. These
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792 Gravel 3. Montreuil, J., Bouque
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794 Gravel
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796 Goodarzi, Fotinopoulou, Turner
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Page 781 and 782:
804 Hounsell, Davies, and Smith 2.
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Monosaccharide Analysis by GC 809 1
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Linkage and Substitution Patterns 8
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Linkage and Substitution Patterns 8
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Page 795 and 796:
820 Hounsell, Davis, and Smith 6. T
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Page 799 and 800:
824 Mizuochi and Hounsell 3. Method
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826 Mizuochi and Hounsell Reference
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834 Weitzhandler et al. alditols (1
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836 Weitzhandler et al. Fig. 1. HPA
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838 Weitzhandler et al. 2. Add 0.1
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Microassay of Protein Glycosylation
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Carbohydrate Electrophoresis 851 12
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Carbohydrate Electrophoresis 853 2.
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Carbohydrate Electrophoresis 855 Pl
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Carbohydrate Electrophoresis 859 Fi
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Carbohydrate Electrophoresis 861 wa
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866 Merry blly less expensive than
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868 Merry 8. Whatman no. 3 chromato
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870 Merry 4. Dialyze for a minimum
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872 Merry 9. Leave for 5 min and un
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874 Merry 2. Column: Reverse-phase
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Table 1 Incubation Conditions for E
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878 Merry 3.7. Exoglycosidase Diges
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880 Merry Fig. 6. Example of exogly
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882 Merry 17. Rice, K. G., Takahash
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Glycoprofiling and SPR 885 124 Glyc
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Glycoprofiling and SPR 887 2. Mater
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Glycoprofiling and SPR 889 Fig. 3.
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Glycoprofiling and SPR 891 α2-3 Ne
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894 Turnbull Fig. 1. Basic principl
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896 Turnbull 13. Enzyme buffer (0.2
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898 Turnbull Table 1 Exoenzymes for
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900 Turnbull Fig. 3. IGS of a hepar
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902 Turnbull 4. Notes 1. Using larg
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904 Turnbull 20. Rice, K., Rottink,
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906 Hooker and James 3. High-perfor
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908 Hooker and James Fig. 1. Whole
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910 Hooker and James at individual
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912 Hooker and James Fig. 4. Sialyl
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914 Hooker and James 16. Ashton, D.
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Lectin Affinity Chromatography 917
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Page 901 and 902:
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Antibody Production 935 128 Antibod
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Antibody Production 937 1.1. Donor
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Antibody Production 939 2. Material
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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:
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Page 927 and 928:
Page 929 and 930:
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964 Bailey is oxidized by chloramin
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The Lactoperoxidase Method 967 133
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The Bolton and Hunter Method 969 13
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Radioiodination Using IODO-GEN 971
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Page 945 and 946:
980 Bailey 3. Specific antiseum to
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982 Bailey Amount of radioactivity
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984 Page and Thorpe 2. Centrifuge s
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DEAE-Sepharose Chromatography 987 1
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IgG Purification with Ion-Exchange
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PEG Precipitation 991 141 Purificat
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994 Page and Thorpe 2. Materials 1.
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996 Dolman and Thorpe Fig. 1. Typic
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Antigen-Ligand Columns 999 144 Puri
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Antigen-Ligand Columns 1001 4. When
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1004 Page and Thorpe 7. Filter and
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1006 Page and Thorpe Fig. 1. SDS-PA
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Purification of IgY from Chicken Eg
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Purification of IgY from Chicken Eg
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1014 Fassina et al. limit the use o
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1016 Fassina et al. 2. Materials Ta
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1018 Fassina et al. 10. Filter the
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1020 Fassina et al. 1. Dilute sampl
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1022 Fassina et al. analysis indica
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1024 Fassina et al. 6. Khayam-Bashi
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1026 Hammerl et al. down, with the
Page 987 and 988:
1028 Hammerl et al. 1. Clean glass
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1030 Hammerl et al. Fig. 1. Experim
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1032 Hammerl et al. can “corrode
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Single-Chain Antibodies 1035 150 Ba
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Single-Chain Antibodies 1037 ing th
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Single-Chain Antibodies 1039 6. Mag
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Single-Chain Antibodies 1041 XL1-Bl
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Single-Chain Antibodies 1043 fore,
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Single-Chain Antibodies 1045 16. Ho
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Enzymatic Digestion of MAbs 1047 15
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Enzymatic Digestion of MAbs 1049 2.
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Enzymatic Digestion of MAbs 1051 2.
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Making Bispecific Antibodies 1053 1
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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-
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Phage Display 1063 11. Agarose top:
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Phage Display 1065 3.1.3.2. AFFINIT
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Phage Display 1067 8. Shake vigorou
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Phage Display 1069 4. Notes 1. Fila
Page 1029 and 1030:
Phage Display 1071 9. Sengupta J.,
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Selection by Panning 1073 154 Scree
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Selection by Panning 1075 Fig. 1. F
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Selection by Panning 1077 12. Centr
Page 1037 and 1038:
Selection by Panning 1079 4. Notes
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Selection by Panning 1081 33. The
Page 1041 and 1042:
Antigen Measurement Using ELISA 108
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Enhanced Chemiluminescence 1089 156
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Enhanced Chemiluminescence 1091 Tab
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Enhanced Chemiluminescence 1093 is
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Enhanced Chemiluminescence 1095 lig
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Immunoprecipitation 1097 157 Immuno
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Immunoprecipitation 1099 Table 2 Pr
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Immunoprecipitation 1101 oligosacch
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Immunoprecipitation 1103 6. Very ge
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Immunoprecipitation 1105 2. Dry the
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Short Chapter Title 1107 PART VIII
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1110 Page and Thorpe final quantity
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1112 Page and Thorpe 2. Materials 1
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Page 1073 and 1074:
161 From: The Protein Protocols Han
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162 From: The Protein Protocols Han
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Affinity Purification of Monoclonal
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Page 1083 and 1084:
Page 1085 and 1086:
An Efficient Method for MAb Product
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Index 1139 Index A Absorbance (UV),
Page 1097 and 1098:
Index 1141 Electrophoresis of antib
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Index 1143 of glycoproteins, 905-91
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Index 1145 India ink, 338 lectin st
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The Protein Protocols Handbook Seco
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