Antibody Phage Display Methods and Protocols - part 2 pot

... in H 2 O. 14. DNase I (RNase-free) and manufacturer’s 10X buffer. 15. PhenolϺchloroformϺisoamyl alcohol (25 Ϻ 24 Ϻ1). 2. 2. RT and PCR Reactions 1. 10X PCR reaction buffer (commercial). 2. 25 ... J. Biol. Chem. 26 8, 53 02 5308. 85. He, M. and Taussig, M. J. (1997) Antibody- ribosome-mRNA (ARM) complexes as effi cient selection particles for in vitro display and evoluti...

Ngày tải lên: 10/08/2014, 00:21

... with complementary methods to increase the scope and range of applications of this technology. 2. Antibody Phage Display 2. 1. The Phage- Display Principle The power of the phage- display system is ... Biology, vol. 178: Antibody Phage Display: Methods and Protocols Edited by: P. M. O’Brien and R. Aitken © Humana Press Inc., Totowa, NJ 1 Overview of Antibody...

Ngày tải lên: 10/08/2014, 00:21

... CC-3′ HuV λ 3B-Back-APA 5′-ACC GCC TCC ACC AGT GCA CTT TCT TCT GAG CTG ACT CAG GAC CC-3′ HuV λ 4-Back-APA 5′-ACC GCC TCC ACC AGT GCA CAC GTT ATA CTG ACT CAA CCG CC-3′ HuV λ 5-Back-APA 5′-ACC ... TC-3′ HuV λ 6-Back-APA 5′-ACC GCC TCC ACC AGT GCA CTT AAT TTT ATG CTG ACT CAG CCC CA-3′ HuV λ 7/8-Back-APA 5′-ACC GCC TCC ACC AGT GCA CAG RCT GTG GTG ACY CAG GAG CC-3′ HuV λ 9-Back-APA 5′-ACC ......

Ngày tải lên: 10/08/2014, 00:21

... ACC 133 From: Methods in Molecular Biology, vol. 178: Antibody Phage Display: Methods and Protocols Edited by: P. M. O’Brien and R. Aitken © Humana Press Inc., Totowa, NJ 9 Panning of Antibody Phage- Display ... solution. Panning Ab Phage- Display Libraries 141 147 From: Methods in Molecular Biology, vol. 178: Antibody Phage Display: Methods and Protocols E...

Ngày tải lên: 10/08/2014, 00:21

... 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 ... (5) and (6) unbound free and labeled hapten removed by washing. 187 From: Methods in Molecular Biology, vol. 178: Antibody Phage Display: Methods and Protocols Edited by...

Ngày tải lên: 10/08/2014, 00:21

... 0.5. Cell Surface Antigen-Binding scFvs 22 9 20 7 From: Methods in Molecular Biology, vol. 178: Antibody Phage Display: Methods and Protocols Edited by: P. M. O’Brien and R. Aitken © Humana Press ... titered (see Note 2) . 2. Human tumor cell lines for selection and absorption, e.g., melanoma cell line, SK-MEL -2 8 , and B-lymphoid cell line, LG -2 . Cell lines should...

Ngày tải lên: 10/08/2014, 00:21

... 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, ... immunohistochemistry and FACS (see Note 10). 24 2 Radoˇsevi´c and van Ewijk 25 5 From: Methods in Molecular Biology, vol. 178: Antibody Phage Display:...

Ngày tải lên: 10/08/2014, 00:21

... 0 .22 0 .20 0 .27 1. 82 14 52. 0 <0.1 52. 0 53.6 0.4 <0.1 0 .22 0 .20 0.34 2. 36 15 52. 5 <0.1 52. 5 54.6 0.4 <0.1 0 .23 0 .20 0. 42 2.90 16 53.0 <0.1 53.0 55.5 0.4 <0.1 0 .23 0 .20 0.57 4.00 ... MgCl 2 0.56 0.90 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...

Ngày tải lên: 10/08/2014, 00:21

... 10.8 g CaCl 2 6H 2 O in 20 mL H 2 O. Sterilize by fi ltration through a 0 .22 µmfi lter. Store at 20 °C. 17. 2X HeBs buffer: 1.6 g NaCl; 0.074 g KCl; 0. 027 g Na 2 HPO 4 , 2H 2 O; 0 .2 g dextrose; ... single-stranded DNA, 27 4 ligation, see Ligation quantitation, 50, 56, 66, 75 E EGF, 22 Electroporation, methods, 52, 66, 94, 29 3, 309, 314, 3 52, 363, 365 preparation of cell...

Ngày tải lên: 10/08/2014, 00:21

... Differential Display 7 H-T110 H-T110 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). ... 5 min Run 32 Donohue, Hsu, and Winkles FGF- 1 M 0 ’ 2h 12h ’ - 28 1 /27 1 - 23 4- 194 - ZF/PTK Primers Fig. 2. Identification of an FGF- 1 -inducible mRNA by dif...

Ngày tải lên: 11/04/2014, 09:39