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546 Judd 4. Excise the protein band from the NCP (a 1 × 5-mm band is more than ample), and place the excised strip in a 1.5-mL microfuge tube. Wash with H 2O until no stain is released into the supernatant. The protein is now ready for labeling and cleavage (see Note 4). 3.1.1.2. DOUBLE SDS-PAGE SEPARATION 1. Separate the samples in individual lanes of an SDS-PAGE gel or in “preparative” SDS-PAGE gels. Fix, stain with CBB, and then destain (see Chapter 11). 2. Excise the protein bands of interest. Soak the bands in 50% ethanol–50% stacking buffer (1 M Tris-HCl, pH 6.8) for 30 min to shrink the gel strip to facilitate loading onto a second SDS-PAGE gel. 3. Push the excised band into contact with the stacking gel of a second SDS-PAGE gel of a different acrylamide concentration (generally use high concentration in the first gel and lower concentration in the second gel). 4. Separate proteins in a second gel (CBB runs just behind the dye front). Stain or electroblot the proteins as in Subheading 3.1.1. The protein is now ready for labeling and cleavage. 3.2. <strong>Protein</strong> Labeling <strong>Protein</strong>s can be intrinsically labeled by growing organisms in the presence of a uniform mixture of 14C-amino acids (5), but this is quite expensive. Intrinsic labeling with individual amino acids, such as 35S-Met or 35S-Cys, will not work, since many peptide fragments will not be labeled. Iodination with 125I is inexpensive and reproducible. Iodinated peptides are readily visualized by autoradiography (1,3,4). Comparative cleavages of a 40,000-Dalton protein intrinsically labeled with 14C-amino acids extrinsically labeled with 125I showed that 61 of 66 α-chymotryptic peptides were labeled with 125I, whereas all 22 Staphylococcus aureus V8 protease-generated peptides were labeled with 125I, demonstrating the effectiveness of radioiodination (10). This demonstrates that tyrosine (Tyr) is not the only amino acid labeled using this procedure. Iodination mediated by chloramine-T (CT) (11) produces extremely high specific activities, but the procedure requires an extra step to remove the CT and can cleave some proteins at tryptophan residues (12). This can be beneficial since it is specific and increases the number of peptides, thus increasing the sensitivity of the procedure (see ref. 13) for peptide maps of CT- vs Iodogen-labeled proteins). Unfortunately, small peptides generated by CT cleavage, followed by a second enzymatic or chemical cleavage, can be lost during the removal of the CT and unbound 125I. The 1,3,4,6-tetrachloro-3α,6α-glycouril (Iodogen) (13) procedure, where the oxidizing agent is bound to the reaction vessel, does not damage the protein and produces high specific activities. Aspiration of the reaction mixture stops the iodination and separates the protein from the oxidant in a single step. For these reasons, Iodogenmediated labeling is the preferred method for radioiodination. (Radioemission of 125I will be expressed as counts per minute [cpm]. This assumes a detector efficiency of 70%. If detector efficiency varies, multiply the cpm presented here by 1.43 to determine decays per minute (dpm), and then multiply the dpm by the efficiency of your detector.) 3.2.1. NCP Strip (Preferred Method) 1. Put the protein-containing NCP strip in an Iodogen-coated (10 µg) microfuge tube. 2. Add 50–100 µL PBS, pH 7.4 (any dilute, neutral buffer should work) and 50–100 µCi 125I (as NaI, carrier-free, 25 µCi/µL) (see Notes 1 and 2).
2-D TLE-TLC Mapping 547 3. Incubate at room temperature for 1 h. Aspirate the supernatant. (Caution: supernatant is radioactive). 4. Place the NCP strip in a fresh microfuge tube and wash three to five times with 1.5 mL H 2O (radioactivity released should stabilize at
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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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Page 487 and 488: Alternative conditions for rapid re
Page 489 and 490: Cleavage at Tryptophanyl-x Bonds 49
Page 491 and 492: Cleavage at Aspartyl-X Bonds 499 73
Page 493 and 494: Cleavage at Aspartyl-X Bonds 501 pr
Page 495 and 496: Cleavage at Cysteinyl-x Bonds 503 7
Page 497 and 498: Cleavage at Cysteinyl-x Bonds 505 F
Page 499 and 500: Cleavage at Asparaginyl-Glycyl Bond
Page 501 and 502: Cleavage at Asparaginyl-Glycyl Bond
Page 503 and 504: Enzymatic Digestion 511 76 Enzymati
Page 505 and 506: Enzymatic Digestion 513 then remove
Page 507 and 508: Enzymatic Digestion 515 Fig. 1. In-
Page 509 and 510: Enzymatic Digestion 517 In the case
Page 511 and 512: Enzymatic Digestion 519 Another app
Page 513 and 514: Enzymatic Digestion 521 11. Modific
Page 515 and 516: 524 Table 1 Digestion Buffers Recip
Page 517 and 518: 526 Fernandez and Mische Fig. 1. Pe
Page 519 and 520: 528 Fernandez and Mische 3. Excise
Page 521 and 522: 530 Fernandez and Mische 3. The lar
Page 523 and 524: 532 Fernandez and Mische membrane a
Page 525 and 526: 534 Stone and Williams 2. Materials
Page 527 and 528: 536 Stone and Williams In those few
Page 529 and 530: 538 Stone and Williams (while maint
Page 531 and 532: 540 Stone and Williams Biochemistry
Page 533 and 534: 2-D TLE-TLC Mapping 543 79 Peptide
Page 535: 2-D TLE-TLC Mapping 545 12. 0.25% N
Page 539 and 540: 2-D TLE-TLC Mapping 549 Table 1 Cle
Page 541 and 542: 2-D TLE-TLC Mapping 551 Fig. 1. Exa
Page 543 and 544: SDS-PAGE Mapping 553 80 Peptide Map
Page 545 and 546: SDS-PAGE Mapping 555 3. Overlay sam
Page 547 and 548: SDS-PAGE Mapping 557 Fig. 1. Exampl
Page 549 and 550: 560 Judd Fig. 1. Example of peptide
Page 551 and 552: Protein Hydrolysates 563 82 Product
Page 553 and 554: Protein Hydrolysates 565 2. Pronase
Page 555 and 556: Amino Acid Analysis with Marfey’s
Page 561 and 562: HPSEC 573 84 Molecular Weight Estim
Page 563 and 564: HPSEC 575 Table 1 Protein Standards
Page 565 and 566: HPSEC 577 Fig. 2. Plot of K d vs lo
Page 567 and 568: HPSEC 579 with care (see suppliers
Page 569 and 570: 582 Aitken larly good resolution de
Page 571 and 572: Disulfide-Linked Peptide Detection
Page 573 and 574: Disulfide-Linked Peptide Detection
Page 575 and 576: Disulfide Bridges 589 87 Diagonal E
Page 577 and 578: Disulfide Bridges 591 3. Spot the s
Page 579 and 580: Disulfide Bridges 593 2. The moveme
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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
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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.
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Detection of Glycosylated Proteins
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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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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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820 Hounsell, Davis, and Smith 6. T
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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
Page 869 and 870:
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
Page 909 and 910:
Production of Anitbodies Using Prot
Page 911 and 912:
Production of Anitbodies Using Prot
Page 913 and 914:
Production of Polyclonal Antibodies
Page 915 and 916:
Page 917 and 918:
Page 919 and 920:
Page 921 and 922:
Peptide Conjugation and Immunizatio
Page 923 and 924:
Page 925 and 926:
Page 927 and 928:
Page 929 and 930:
Page 931 and 932:
964 Bailey is oxidized by chloramin
Page 933 and 934:
The Lactoperoxidase Method 967 133
Page 935 and 936:
The Bolton and Hunter Method 969 13
Page 937 and 938:
Radioiodination Using IODO-GEN 971
Page 939 and 940:
Page 941 and 942:
Page 943 and 944:
Page 945 and 946:
980 Bailey 3. Specific antiseum to
Page 947 and 948:
982 Bailey Amount of radioactivity
Page 949 and 950:
984 Page and Thorpe 2. Centrifuge s
Page 951 and 952:
DEAE-Sepharose Chromatography 987 1
Page 953 and 954:
IgG Purification with Ion-Exchange
Page 955 and 956:
PEG Precipitation 991 141 Purificat
Page 957 and 958:
994 Page and Thorpe 2. Materials 1.
Page 959 and 960:
996 Dolman and Thorpe Fig. 1. Typic
Page 961 and 962:
Antigen-Ligand Columns 999 144 Puri
Page 963 and 964:
Antigen-Ligand Columns 1001 4. When
Page 965 and 966:
1004 Page and Thorpe 7. Filter and
Page 967 and 968:
1006 Page and Thorpe Fig. 1. SDS-PA
Page 969 and 970:
Purification of IgY from Chicken Eg
Page 971 and 972:
Purification of IgY from Chicken Eg
Page 973 and 974:
1014 Fassina et al. limit the use o
Page 975 and 976:
1016 Fassina et al. 2. Materials Ta
Page 977 and 978:
1018 Fassina et al. 10. Filter the
Page 979 and 980:
1020 Fassina et al. 1. Dilute sampl
Page 981 and 982:
1022 Fassina et al. analysis indica
Page 983 and 984:
1024 Fassina et al. 6. Khayam-Bashi
Page 985 and 986:
1026 Hammerl et al. down, with the
Page 987 and 988:
1028 Hammerl et al. 1. Clean glass
Page 989 and 990:
1030 Hammerl et al. Fig. 1. Experim
Page 991 and 992:
1032 Hammerl et al. can “corrode
Page 993 and 994:
Single-Chain Antibodies 1035 150 Ba
Page 995 and 996:
Single-Chain Antibodies 1037 ing th
Page 997 and 998:
Single-Chain Antibodies 1039 6. Mag
Page 999 and 1000:
Single-Chain Antibodies 1041 XL1-Bl
Page 1001 and 1002:
Single-Chain Antibodies 1043 fore,
Page 1003 and 1004:
Single-Chain Antibodies 1045 16. Ho
Page 1005 and 1006:
Enzymatic Digestion of MAbs 1047 15
Page 1007 and 1008:
Enzymatic Digestion of MAbs 1049 2.
Page 1009 and 1010:
Enzymatic Digestion of MAbs 1051 2.
Page 1011 and 1012:
Making Bispecific Antibodies 1053 1
Page 1013 and 1014:
Page 1015 and 1016:
Making Bispecific Antibodies 1057 F
Page 1017 and 1018:
Phage Display 1059 153 Phage Displa
Page 1019 and 1020:
Phage Display 1061 5000, 1 mL of 2-
Page 1021 and 1022:
Phage Display 1063 11. Agarose top:
Page 1023 and 1024:
Phage Display 1065 3.1.3.2. AFFINIT
Page 1025 and 1026:
Phage Display 1067 8. Shake vigorou
Page 1027 and 1028:
Phage Display 1069 4. Notes 1. Fila
Page 1029 and 1030:
Phage Display 1071 9. Sengupta J.,
Page 1031 and 1032:
Selection by Panning 1073 154 Scree
Page 1033 and 1034:
Selection by Panning 1075 Fig. 1. F
Page 1035 and 1036:
Selection by Panning 1077 12. Centr
Page 1037 and 1038:
Selection by Panning 1079 4. Notes
Page 1039 and 1040:
Selection by Panning 1081 33. The
Page 1041 and 1042:
Antigen Measurement Using ELISA 108
Page 1043 and 1044:
Page 1045 and 1046:
Page 1047 and 1048:
Enhanced Chemiluminescence 1089 156
Page 1049 and 1050:
Enhanced Chemiluminescence 1091 Tab
Page 1051 and 1052:
Enhanced Chemiluminescence 1093 is
Page 1053 and 1054:
Enhanced Chemiluminescence 1095 lig
Page 1055 and 1056:
Immunoprecipitation 1097 157 Immuno
Page 1057 and 1058:
Immunoprecipitation 1099 Table 2 Pr
Page 1059 and 1060:
Immunoprecipitation 1101 oligosacch
Page 1061 and 1062:
Immunoprecipitation 1103 6. Very ge
Page 1063 and 1064:
Immunoprecipitation 1105 2. Dry the
Page 1065 and 1066:
Short Chapter Title 1107 PART VIII
Page 1067 and 1068:
1110 Page and Thorpe final quantity
Page 1069 and 1070:
1112 Page and Thorpe 2. Materials 1
Page 1071 and 1072:
Page 1073 and 1074:
161 From: The Protein Protocols Han
Page 1075 and 1076:
162 From: The Protein Protocols Han
Page 1077 and 1078:
Affinity Purification of Monoclonal
Page 1079 and 1080:
Page 1081 and 1082:
Page 1083 and 1084:
Page 1085 and 1086:
An Efficient Method for MAb Product
Page 1087 and 1088:
Page 1089 and 1090:
Page 1091 and 1092:
Page 1093 and 1094:
Page 1095 and 1096:
Index 1139 Index A Absorbance (UV),
Page 1097 and 1098:
Index 1141 Electrophoresis of antib
Page 1099 and 1100:
Index 1143 of glycoproteins, 905-91
Page 1101 and 1102:
Index 1145 India ink, 338 lectin st
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The Protein Protocols Handbook Seco
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