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122 Waterborg and quantitate, intensity of protein staining patterns. Care should be taken to develop a standard scanning setup, using the full dynamic range (typically all three colors used at their full range, 0–255 for a 24-bit scanner; excluding automatic adjustments for density and contrast). A standard gamma correction value should be determined, using an optical density wedge (Kodak), to assure that the density response is linear. Placing a destained polyacrylamide gel on the gel scanner, one should cover the top of the gel with an acetate film to prevent surface reflection abnormalities and prevent touching of the transilluminating light source surface to the film, avoiding moiré interference patterns. Recording gel patterns at 300 dpi in full color and saving loss-less tiff files facilitates faithful replication of experimental results (Fig. 1). Quantitative densitometry programs can use the image files. 14. Triton X-100 interferes with native-mode electrotransfer of histones to nitrocellulose (15). Exchange of Triton by SDS under acidic conditions allows Western blotting (16) (see Chapter 42). References 1. Zweidler, A. (1978) Resolution of histones by polyacrylamide gel electrophoresis in presence of nonionic detergents. Meth. Cell Biol. 17, 223–233. 2. Manca, L., Cherchi, L., De Rosa, M. C., Giardina, B., and Masala, B. (2000) A new, electrophoretically silent, fetal hemoglobin variant: Hb F-Calabria [Ggamma118 (GH1) Phe→Leu]. Hemoglobin 24, 37–44. 3. Urban, M. K., Franklin, S. G., and Zweidler, A. (1979) Isolation and characterization of the histone variants in chicken erythrocytes. Biochemistry 18, 3952–3960. 4. Schwager, S. L. U., Brandt, W. F., and Von Holt, C. (1983) The isolation of isohistones by preparative gel electrophoresis from embryos of the sea urchin Parechinus angulosus. Biochim. Biophys. Acta 741, 315–321. 5. Waterborg, J. H., Harrington, R. E., and Winicov, I. (1987) Histone variants and acetylated species from the alfalfa plant Medicago sativa. Arch. Biochem. Biophys. 256, 167–178. 6. Bonner, W. M., West, M. H. P., and Stedman, J. D. (1980) Two-dimensional gel analysis of histones in acid extracts of nuclei, cells, and tissues. Eur. J. Biochem. 109, 17–23. 7. Waterborg, J. H. (1992) Existence of two histone H3 variants in dicotyledonous plants and correlation between their acetylation and plant genome size. Plant Mol. Biol. 18, 181–187. 8. Waterborg, J. H. (1991) Multiplicity of histone H3 variants in wheat, barley, rice and maize. Plant Physiol. 96, 453–458. 9. Waterborg, J. H., Robertson, A. J., Tatar, D. L., Borza, C. M., and Davie, J. R. (1995) Histones of Chlamydomonas reinhardtii. Synthesis, acetylation and methylation. Plant Physiol. 109, 393–407. 10. Waterborg, J. H. (2000) Steady-state levels of histone acetylation in Saccharomyces cerevisiae. J. Biol. Chem. 275, 13,007–13,011. 11. Waterborg, J. H. and Matthews, H. R. (1983) Patterns of histone acetylation in the cell cycle of Physarum polycephalum. Biochemistry 22, 1489–1496. 12. Arents, G., Burlingame, R. W., Wang, B. C., Love, W. E., and Moudrianakis, E. N. (1991) The nucleosomal core histone octamer at 3.1 Å resolution. A tripartite protein assembly and a left-handed superhelix. Proc. Natl. Acad. Sci. USA 88, 10,138–10,148. 13. Luger, K., Mäder, A. W., Richmond, R. K., Sargent, D. F., and Richmond, T. J. (1997) Crystal structure of the nucleosome core particle at 2.8Å resolution. Nature 389, 251–260. 14. Sullivan, S. A., Aravind, L., Makalowska, I., Baxevanis, A. D., and Landsman, D. (2000) The histone database: a comprehensive WWW resource for histones and histone fold-containing proteins. Nucleic Acids Res. 28, 320–322.
AUT Gel for Histones 123 15. Waterborg, J. H. and Harrington, R. E. (1987) Western blotting from acid–urea–Triton– and sodium dodecyl sulfate-polyacrylamide gels. Analyt. Biochem. 162, 430–434. 16. Delcuve, G. P. and Davie, J. R. (1992) Western blotting and immunochemical detection of histones electrophoretically resolved on acid–urea–Triton- and sodium dodecyl sulfatepolyacrylamide gels. Analyt. Biochem. 200, 339–341.
Page 1 and 2:
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
Page 85 and 86: SDS-PAGE 67 close to the dye, and t
Page 87 and 88: 70 Walker 2. Buffers: a. 1.875 M Tr
Page 89 and 90: 72 Walker Fig. 2. Diagrammatic repr
Page 91 and 92: 74 Judd 2. Materials 2.1. Equipment
Page 93 and 94: 76 Judd ber with anode buffer. Remo
Page 95 and 96: 78 Judd 4. Run the gel as described
Page 97 and 98: SDecS-PAGE of Nucleic Acid Binding
Page 103 and 104: CAT Gel Electrophoresis 87 15 Cetyl
Page 105 and 106: CAT Gel Electrophoresis 89 Fig. 1.
Page 107 and 108: CAT Gel Electrophoresis 91 3. CAT s
Page 109 and 110: CAT Gel Electrophoresis 93 Table 1
Page 111 and 112: CAT Gel Electrophoresis 95 the tank
Page 113 and 114: CAT Gel Electrophoresis 97 may be a
Page 115 and 116: CAT Gel Electrophoresis 99 enzyme p
Page 117 and 118: CAT Gel Electrophoresis 101 20. Rey
Page 119 and 120: 104 Waterborg Fig. 1. Histones of t
Page 121 and 122: 106 Waterborg 9. Riboflavin-5'-phos
Page 123 and 124: 108 Waterborg 25. Remove the bottom
Page 125 and 126: 110 Waterborg 10. The amount of pro
Page 127 and 128: AUT Gel for Histones 113 17 Acid-Ur
Page 129 and 130: AUT Gel for Histones 115 Details of
Page 131 and 132: AUT Gel for Histones 117 Fig. 2. (A
Page 133 and 134: AUT Gel for Histones 119 13. Add 0.
Page 135: AUT Gel for Histones 121 5. The sep
Page 139 and 140: 126 Walker the cost of preparing IE
Page 141 and 142: 128 Walker 4. Notes 1. The sucrose
Page 143 and 144: Protein Solubility in 2-D Electroph
Page 153 and 154: Fractionated Extraction 141 20 Prep
Page 155 and 156: Fractionated Extraction 143 Fig. 1.
Page 157 and 158: Fractionated Extraction 145 Table 2
Page 159 and 160: Fractionated Extraction 147 13. Com
Page 161 and 162: 149 SI + II fraction: liver Brain H
Page 163 and 164: Fractionated Extraction 151 3. Add
Page 165 and 166: Fractionated Extraction 153 Fig. 2.
Page 167 and 168: Fractionated Extraction 155 and bra
Page 169 and 170: Fractionated Extraction 157 ume are
Page 171 and 172: 160 Bizios 2. Materials 2.1. Equipm
Page 173 and 174: 162 Bizios 5. Sample preparation sh
Page 175 and 176: 164 Gravel Fig. 1. Tube Cell Model
Page 177 and 178: 166 Gravel 3. Fill the lower chambe
Page 179 and 180: 168 Gravel lysis solution A (SDS-DT
Page 181 and 182: 170 Gianazza Fig. 1. Structure of t
Page 183 and 184: 172 Gianazza Table 1 pK Values of I
Page 185 and 186: 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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Immunoblotting of 2-DE Separated Pr
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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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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
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252 Fernandez-Patron to years witho
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254 Fernandez-Patron Fig. 2. Revers
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256 Fernandez-Patron spectrometry (
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258 Fernandez-Patron 11. Fernandez-
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260 Choi, Hong, and Yoo Table 1 Lin
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262 Choi, Hong, and Yoo Fig. 2. Mec
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Silver Staining of Proteins 265 33
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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.
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274 Patton plexes for colorimetric
Page 281 and 282:
276 Patton device (CCD) camera. The
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278 Patton 3.2. Luminescent Detecti
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280 Patton 3.5. Luminescent Detecti
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282 Patton EDTA, pH 9.6 or using SY
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284 Patton filter. Proteins stained
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286 Patton 17. Lim, M., Patton, W.,
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288 Dunn variations in silver stain
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290 Dunn Fig. 1. A 2-DE separation
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292 Dunn 11. Stephens, R. E. (1975)
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Gels Using Eosin Y Stain 295 36 Det
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Gels Using Eosin Y Stain 297 3. An
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300 Jenö and Horst and Coomassie B
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302 Jenö and Horst Fig. 1. Side (A
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304 Jenö and Horst BT1 membrane, o
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Autoradiography and Fluorography 30
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Page 313 and 314:
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Page 317 and 318:
Blotting-Electroblotting 315 PART I
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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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Page 342 and 343:
Page 344 and 345:
344 Wisdom 4. Glutaraldehyde. 5. 50
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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
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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
Page 364 and 365:
Preparation of Avidin Conjugates 36
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Page 372 and 373:
Page 374 and 375:
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.
Page 380 and 381:
382 Root and Wang suspension become
Page 382 and 383:
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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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
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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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Page 445 and 446:
Page 447 and 448:
Page 449 and 450:
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
Page 455 and 456:
460 Aitken and Learmonth 4. Notes 1
Page 457 and 458:
462 Ward Fig. 1. Amino acid sequenc
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Chemical Modifications of Proteins
Page 461 and 462:
Chemical Modifications of Proteins
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470 Tawfik 3. Method 3.1. Nitration
Page 465 and 466:
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
Page 519 and 520:
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
Page 541 and 542:
2-D TLE-TLC Mapping 551 Fig. 1. Exa
Page 543 and 544:
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
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
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HPSEC 579 with care (see suppliers
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582 Aitken larly good resolution de
Page 571 and 572:
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
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
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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
Page 609 and 610:
624 Weber and McFadden Fig. 1. Sche
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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
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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
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650 Corsini Fig. 5. Metabolic label
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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
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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
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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.
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Phosphopeptide Mapping 679 until th
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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
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Nanoelectrospray MS/MS for Peptide
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Page 691 and 692:
Page 693 and 694:
MALDI-MS for Protein Identification
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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
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744 Hennig In the age of genomics,
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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
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Linkage and Substitution Patterns 8
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Linkage and Substitution Patterns 8
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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
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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
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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
Page 871 and 872:
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
Page 881 and 882:
910 Hooker and James at individual
Page 883 and 884:
912 Hooker and James Fig. 4. Sialyl
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914 Hooker and James 16. Ashton, D.
Page 887 and 888:
Lectin Affinity Chromatography 917
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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
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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:
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
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The Bolton and Hunter Method 969 13
Page 937 and 938:
Radioiodination Using IODO-GEN 971
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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
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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
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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
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Page 1045 and 1046:
Page 1047 and 1048:
Enhanced Chemiluminescence 1089 156
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Enhanced Chemiluminescence 1091 Tab
Page 1051 and 1052:
Enhanced Chemiluminescence 1093 is
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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
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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
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Index 1145 India ink, 338 lectin st
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The Protein Protocols Handbook Seco
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