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Short Chapter Title 819 114 O-Linked Oligosaccharide Profiling by HPLC Elizabeth F. Hounsell, Michael J. Davies, and Kevin D. Smith 1. Introduction The several different core regions of O-linked chains (Chapter 113) can be further extended by Gal and GlcNAc containing backbones or by addition of blood group antigen-type glycosylation and the presence of sialic acid or sulfate. We sought a universal column that can be applied with high resolution to the various oligosaccharide alditols released from glycoproteins by β-elimination (see Note 1) and have pioneered (1,2) the use of porous graphitized carbon (PGC). This is an alternative to C 18 reversedphase HPLC and normal-phase amino-bonded columns, which can be used together in the presence and absence of high-salt buffers (3). 2. Materials 1. Gradient HPLC system: e.g., 2 × 302 pumps, 802C manometric module, 811 dynamic mixer, 116 UV detector, 201 fraction collector, 715 chromatography system control software (all Gilson Medical Electronics, Villiers-le-Bel, France). 2. IBM PS-2 personal computer (or compatible model) with Microsoft Windows. 3. Hypercarb S HPLC Column (100 × 4.6 mm) (Shandon Scientific, Runcorn, Cheshire, England) or Glycosep H (100 × 3 mm, Oxford Glycosystems, Abingdon, UK). 4. HPLC-grade H2O. 5. HPLC-grade acetonitrile. 6. HPLC-grade trifluoroacetic acid (Pierce and Warriner, Chester, UK). 3. Methods 1. Prepare eluant A: 500 mL of 0.05% TFA. 2. Prepare eluant B: 250 mL of acetonitrile containing 0.05% TFA. 3. Degas eluants by sparging with helium. 4. Equilibrate the column in 100% eluant A and 0% eluant B for 30 min at 0.75 mL/min prior to injection samples. 5. Elute 10 nmol oligosaccharide alditol (obtained as in Chapter 113) with the following gradient at a flow rate of 0.75 mL/min and UV detection at 206 nm/0.08 AUFS. a. Time = 0 min A = 100%; B = 0% b. Time = 5 min A = 100%; B = 0% c. Time = 40 min A = 60%; B = 40% d. Time = 45 min A = 60%; B = 40% e. Time = 50 min A = 100%; B = 0% From: The <strong>Protein</strong> <strong>Protocols</strong> Handbook, 2nd Edition Edited by: J. M. Walker © Humana Press Inc., Totowa, NJ 819
820 Hounsell, Davis, and Smith 6. The resulting oligosaccharide containing fractions are then derivatized for LSIMS and GC-MS or analyzed by NMR. 4. Note 1. The GalNAc residue linked to Ser/Thr in O-linked chains is normally substituted at least at C-3 and therefore the alkali catalyzed β-elimination reaction will also result in “peeling” of the released oligosaccharide. This is obviated by concomitant reduction to give oligosaccharide alditols ending in GalNAcol. Endo-α-N-acetylgalactosaminidase digestion at hydrazinolysis under mild conditions can be used to release intact reducing sugars, which will have longer retention times on HPLC (4). References 1. Davies, M. J., Smith, K. D., Harbin, A.-M., and Hounsell, E. F. (1992) High performance liquid chromatography of oligosaccharide alditols and glycopeptides on graphitized carbon column. J. Chromatogr. 609, 125–131. 2. Davies, M. J., Smith, K. D., Carruthers, R. A., Chai, W., Lawson, A. M., and Hounsell, E. F. (1993) The use of a porous graphitized carbon (PGC) column for the HPLC of oligosaccharides, alditols and glycopeptides with subsequent mass spectrometry analysis. J. Chromatogr. 646, 317–326. 3. Davies, M. J. and Hounsell, E. F. (1996) Comparison of separation modes for high performance liquid chromatography of glycoprotein- and proteoglycan-derived oligosaccharides. J. Chromatogr. 720, 227–234. 4. Hounsel, E. F., ed. (1993) Methods in Molecular Biology, vol 14: Glycoprotein Analysis in Biomedicine, Humana, Totowa, NJ.
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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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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
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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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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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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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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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Glycoprotein Detection 761 104 Dete
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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 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 781 and 782: 804 Hounsell, Davies, and Smith 2.
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 793: 818 Hounsell, Davies, and Smith 3.
Page 799 and 800: 824 Mizuochi and Hounsell 3. Method
Page 801 and 802: 826 Mizuochi and Hounsell Reference
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 825 and 826: Carbohydrate Electrophoresis 851 12
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Page 829 and 830: Carbohydrate Electrophoresis 855 Pl
Page 833 and 834: Carbohydrate Electrophoresis 859 Fi
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Page 839 and 840: 866 Merry blly less expensive than
Page 841 and 842: 868 Merry 8. Whatman no. 3 chromato
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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 893 and 894:
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Page 897 and 898:
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Page 901 and 902:
Page 903 and 904:
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:
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 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
Page 965 and 966:
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
Page 973 and 974:
1014 Fassina et al. limit the use o
Page 975 and 976:
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
Page 993 and 994:
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
Page 999 and 1000:
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.
Page 1009 and 1010:
Enzymatic Digestion of MAbs 1051 2.
Page 1011 and 1012:
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-
Page 1021 and 1022:
Phage Display 1063 11. Agarose top:
Page 1023 and 1024:
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
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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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
Page 1059 and 1060:
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
Page 1069 and 1070:
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
Page 1079 and 1080:
Page 1081 and 1082:
Page 1083 and 1084:
Page 1085 and 1086:
An Efficient Method for MAb Product
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Page 1091 and 1092:
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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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