Antibody Phage Display Methods and Protocols - part 1 docx

... Biology, vol. 17 8: Antibody Phage Display: Methods and Protocols Edited by: P. M. O’Brien and R. Aitken © Humana Press Inc., Totowa, NJ 1 Overview of Antibody Phage- Display Technology and Its Applications Hennie ... with complementary methods to increase the scope and range of applications of this technology. 2. Antibody Phage Display 2 .1. The Phage- Di...

Ngày tải lên: 10/08/2014, 00:21

... may be used. 12 . AP substrate, e.g., combine 0.4 mM 2,2′-di-p-nitrophenyl-5,5′,-diphenyl-3, 3 -[ 3,3′-dimethoxy-4,4′-diphenylene] ditetrazolium chloride and 0.38 mM 5-bromo-4-chloro-3-indoxyl phosphate ... containing 10 mM DTT (fraction 2b). 11 . For SDS-PAGE measurements, add 10 µL 10 X reducing loading buffer to fractions 0 and 1, and load samples (e.g., 10 µL and 50 µL...

Ngày tải lên: 10/08/2014, 00:21

... Titer f (%) g γ 1 κ Fab /phage library panning results 0 0 0/24 (0) 1 2.94 × 10 11 6.04 × 10 5 11 12 .1 1 /16 0 /16 (0) 2 2 .15 × 10 11 1. 68 × 10 7 11 78.3 13 8.0× 1/ 2048 15 /15 (10 0) 3 1. 72 × 10 11 1. 44 × 10 8 18 40.0 ... 1. 44 × 10 8 18 40.0 11 0.7× 1/ 2048 12 /12 (10 0) γ 1 λ Fab /phage library panning results 0 0 0/20 (0) 1 2.28 × 1...

Ngày tải lên: 10/08/2014, 00:21

... 0.20 1. 40 1. 5 50.7 <0 .1 50.7 51. 1 0.4 <0 .1 0.35 0.40 0.20 1. 35 12 51. 0 <0 .1 51. 0 51. 6 0.4 <0 .1 0.20 0.20 0 .18 1. 26 13 51. 5 <0 .1 51. 5 52.6 0.4 <0 .1 0.22 0.20 0.27 1. 82 14 52.0 ... (µL) (µL) 10 X Synthesis buffer 11 .3 11 .3 T4 DNA ligase (3 U/µL) 11 .0 11 .0 T7 DNA polymerase 11 .0 11 .0 (1U/µL diluted 1 1 in T7 polymerase buffer) To...

Ngày tải lên: 10/08/2014, 00:21

... Appropriate phagemid vector (e.g., MC03) (Fig. 1) . 9. Low-melting-temperature agarose. 10 . Long-wave, hand-held UV lamp. 11 . UV-transparent shrink-wrap fi lm. 12 . Scalpel blades. 13 . Ethidium ... transition-state analog by phage display. Appl. Biochem. Biotechnol. 47, 15 7 17 1. 12 7. Goldberg, M. E. and Djavadi, O. L. (19 93) Methods for measurement of antibody/ an...

Ngày tải lên: 10/08/2014, 00:21

... CC-3′ V λ Back HuV λ 1A-Back-APA 5′-ACC GCC TCC ACC AGT GCA CAG TCT GTG CTG ACT CAG CCA CC-3′ HuV λ 1B-Back-APA 5′-ACC GCC TCC ACC AGT GCA CAG TCT GTG YTG ACG CAG CCG CC-3′ HuV λ 1C-Back-APA ... I., Inoue, R., and Ikeda-Tanaka, O. (19 96) Chicken monoclonal antibody isolated by a phage display system. J. Immunol. 15 7, 11 56 11 62. 10 . Cary, S. P., Lee, J., Wagenknecht, R., a...

Ngày tải lên: 10/08/2014, 00:21

... solution. Panning Ab Phage- Display Libraries 14 1 14 7 From: Methods in Molecular Biology, vol. 17 8: Antibody Phage Display: Methods and Protocols Edited by: P. M. O’Brien and R. Aitken © Humana ... isopropyl- - D-thiogalactopyranoside. 9. Bacterial strains: XL1 Blue: recA1, endA1, gyrA 46, thi -1 , hsdR 17 , supE 44, relA1, lac–, (F′ proA + B + , lacI q , lacZ ∆...

Ngày tải lên: 10/08/2014, 00:21

... Mark nine autoclaved 1. 8-mL microcentrifuge tubes as follows: TG 1- 1 0 1 , TG 1- 10 –2 , TG 1- 1 0 –3 , TG 1- 1 0 –4 , CJ23 6 -1 0 –4 , CJ23 6 -1 0 –5 , CJ23 6 -1 0 –6 , CJ236 10 –7 , and CJ23 6 -1 0 –8 . Into each, ... 5 10 pg ssuDNA. Prepare tubes as in Table 1. Table 1 Annealing of Oligo to Template Experimental Control (µL) (µL) H 2 O 17 18 10 X Annealing buf...

Ngày tải lên: 10/08/2014, 00:21

... 224 Carcinoembryonic antigen, 6, 17 , 18 CC-CKR5, 18 CCR5, 368 CD3, 319 CD19, 319 CD28, 319 CD36, 15 CD40, 12 1 CD55, 17 cDNA, storage, 55 synthesis, 47, 65, 75, 83, 90, 97, 11 7, 12 6, 211 c-erbB-2, 39 Chromatography, immobilized ... human anti-gp120 antibody. Hum. Gene. Ther. 7, 15 15 15 25. 3. Marasco, W. A., LaVecchio, J., and Winkler, A. (19 99) Human anti-HIV -1 ta...

Ngày tải lên: 10/08/2014, 00:21

... Differential Display 7 H-T 110 H-T 110 H-TIIA H-WA H-TIIA H-TIIC H-TllC H-AP3 H-APB H-API H-AP3 H-AP3 H-APl H-AP3 Fig. 2. Differential display using one-base anchored oligo-dT primers (7). ... and 72°C for 30 s with a final extension of 5 min Run 32 Donohue, Hsu, and Winkles FGF- 1 M 0 ’ 2h 12 h ’ - 2 81/ 2 71 - 23 4- 19 4 - ZF/PTK Primers Fig. 2. I...

Ngày tải lên: 11/04/2014, 09:39