Antibody Phage Display Methods and Protocols - part 10 pps

... Dissolve 15 g Bacto-agar in 790 mL distilled water and autoclave. Cool to 60°C and add 100 mL 10X stock of yeast nitrogen base without amino acids, 100 mL 10X glucose stock, and 10 mL 100 X stock(s) ... U/µL) and XbaI restriction enzyme (20 U/µL) with bovine serum albumin (BSA) 100 X and NEBuffer 2 10X (500 mM NaCl, 100 mM Tris-HCl, 100 mM MgCl 2 , 10 mM dithiothreitol,...

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

... 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 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- Display...

Ngày tải lên: 10/08/2014, 00:21

... agarose gels and from solution. 8. 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. ... 53 39 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 2 Stan...

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 ... (F(ab) 2 -specifi c or anti-immunoaffi nity tag (e.g., 9E10 anti-myc) mouse MAb; appropriate horse- radish peroxidase (HRP)-conjugated anti-mouse IgG. For detection...

Ngày tải lên: 10/08/2014, 00:21

... 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 mono-(p-toluidinium) salt in 0.1 M Tris- HCl, ... (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: P....

Ngày tải lên: 10/08/2014, 00:21

... Anti-M13 horseradish peroxidase-conjugated Ab (Pharmacia Biotech cat. no. 2 7-9 41 1-0 1); fl uorescein-isothiocyanate-conjugated anti-sheep Ab; anti-M13 Ab (Pharmacia Biotech cat. no. 2 7-9 41 0-0 1). ... 0.5. Cell Surface Antigen-Binding scFvs 229 207 From: Methods in Molecular Biology, vol. 178: Antibody Phage Display: Methods and Protocols Edited by: P. M. O’Brien...

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) . 242 Radoˇsevi´c and van Ewijk 255 From: Methods in Molecular Biology, vol. 178: Antibody Phage Display:...

Ngày tải lên: 10/08/2014, 00:21

... Molecular Biology, vol. 178: Antibody Phage Display: Methods and Protocols Edited by: P. M. O’Brien and R. Aitken © Humana Press Inc., Totowa, NJ 28 Generation of Bispecifi c and Tandem Diabodies Sergey ... tumor-targeting specifi city (Fig. 1). Smaller Ab-derived molecules include enzymatically produced 50-kDa Fabs and engineered 25-kDa single-chain Fvs (scFvs) consisting of...

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). ... non-transformed cell line Rat 1 and H-ras transformed cell lines rat 1 (ras), T10 1-4 and Al-5 (lanes from left to right, respectively) were compared by differential display using t...

Ngày tải lên: 11/04/2014, 09:39