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Phage Display Rabbit Antibody Library Construction Kit


Phagemid: pCDisplay-4 E. coli TG1 host strain: K12 D(lac-pro), supE, thi, hsdD5/F’[traD36, proAB, lacIq, lacZDM15]. TOP10F′host strain: F′{lac lq Tn10 (tetR)}, mcrA, Δ(mrr-hsdRMS-mcrBC), φ80lacZΔM15, ΔlacX74, deoR, recA1, araD139, Δ(ara -leu )7697, galU, galK, rpsL (strr), endA1, nupG M13K07 helper phage: ~1.0×1011pfu phages (~1.0×1012 pfu/mL) supplied in 100μL TBS with 15% glycerol. Kanamycin resistant. The sequencing primers:
Kit storage: In the kit, there are two bacterial strains in stab agar to be stored at 4 °C.  The helper phage should be kept at -20°C. There are two DNA primers to be stored at -20°C as well.


Principle of phage display

In a phage display library, a variety of peptides, small antibodies [e.g. scFv and Fab] or proteins are displayed on the surface of filamentous phage [M13, fd, and f1 strains] or lambda phage as fusion proteins with one of the coat proteins of the phage virions, while the genetic materials encoding the peptides/proteins are housed within the virion. Using a binding-based process called bio-panning [Figure 1], a small number of phages that display proteins specifically binding to a target of interest can be rescued from a phage library that usually displays a repertoire of many billions of unique peptides/proteins. Finally, the peptides/proteins displayed by the selected phages can be identified by phage amplification and DNA sequencing.

Principle of phage display


Media and solution
LB Medium: Per liter: 10 g Bacto-Tryptone, 5 g yeast extract, 5 g NaCl. Autoclave 20 minutes at 121°C.
LB Plates: LB medium + 15 g/L agarose.
Top agar: as for LB medium, but use 7 g/L Bacto-agar. Before use, melt and cool to 50°C.
SB (Super Broth) Medium: 10 g of MOPS, 30 g of tryptone, 20 g of yeast extract, 1 liter volume with ddH2O. Stir to dissolve, titrate to pH 7.0. Autoclave at 121°C for 20 minutes.
PBS: Per liter: 8 g NaCl, 0.2 g KCl, 1.7 g Na2HPO4, 0.163 g KH2PO4, pH to 7.4 with HCl.
Blocking buffer: 0.1M NaHCO3 (pH 8.6), 5 mg/mL BSA, 0.02% NaN3, Filter sterilize, store at 4°C.
Coating Buffer: 0.1M NaHCO3 (pH 8.6).
Conjugated second antibody: 

Human Immunoglobulin G (IgG) Fab'2 Rabbit anti-Human Polyclonal (HRP) Antibody: LifeSpan BioSciences
EIA/RIA strip-well 96-well plates: Corning, USA
Substrate: TMB and H2O2 from Pierce.
5*PEG/NaCl Dissolve in water: 200 g of polyethylene glycol-8000, 150 g of NaCl(2.5M). Bring volume to 1 liter; stir until dissolved. Filter through a 0.8 µm filter.
SOB Medium: To 950 mL of deionized water, add 20 g of tryptone, 5.0 g of yeast extract, 0.5 g of NaCl, 186 mg of KCl. Mix the solution until dissolved, and adjust the pH to 7.0 with NaOH. Adjust the volume to 1 liter. Sterilize by autoclaving for 20 minutes at 15 psi on liquid cycle. When cooled, add 10 mL of sterile 1 M MgCl2.

SOC Medium: Add 1 mL of 1 M glucose (filter-sterilized) to 50 mL of SOB (final = 20 mM).



  1. C. F. Barbas III, D. R. Burton, J. K. Scott, and G. J. Silverman, (2001) Phage Display: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
  2. O'Brien P.M. Aitken R. Totowa, (2001) Antibody phage display: methods and protocols. In: Methods in Molecular Biology, Vol 178. Humana Press, NJ.
  3. Sachdev S. Sidhu, (2005) Phage Display in Biotechnology and Drug Discovery. CRC Press/Taylor & Francis Group, Boca Raton.

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