 | | | | Ph.D.™-12 Phage Display Peptide Library | | | |
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Prices are in US dollars and valid only for US orders.
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Description:
The Ph.D.™-12 Phage Display Peptide Library is based on a combinatorial library of random dodecapeptides fused to a minor coat protein (pIII) of M13 phage (1-6). The displayed peptide (12-mer) is expressed at the N-terminus of pIII, i.e., the first residue of the mature protein is the first randomized position. The peptide is followed by a short spacer (Gly-Gly-Gly-Ser) and then the wild-type pIII sequence. The library consists of approximately 2.7 x 109 electroporated sequences amplified once to yield approximately 100 copies of each sequence in 10 µl of the supplied phage.
Reaction & Storage Conditions
Concentration:
1 x1013 pfu/ml
Storage Conditions:
50% Glycerol
Storage Temperature:
-20°C
Notes
General notes:- Supplied in: TBS with 50% glycerol.
Usage notes:- Complexity: 2.7 x 109 transformants.
FAQs
- Where can I find many more detailed FAQs for Phage Display Peptide Libraries?
- Can a different bacterial strain be used with the Ph.D.™ Phage Display?
- No plaques are visible when titering using the Ph.D.™ Phage Display kit.
- I am using Ph.D.™ Phage display and the amplified phage titer is low.
- I am using Ph.D.™ Phage Display and the phage DNA templates do not yield readable sequence.
- I am using Ph.D.™ Phage Display and the sequencing templates do not run where they should on a gel.
- I am using Ph.D.™ Phage Display and after 4 or more rounds of panning all clones are wild-type phage (white plaques).
- When performing an experiment using Ph.D.™ Phage Display, the ELISA indicates that background binding to the plate is as high as binding to the target.
- When using the Ph.D.™ Phage Display, panning yielded a consensus sequence, but no ELISA signal.
- I am using Ph.D.™ Phage Display and the streptavidin control experiment did not yield the HPQ consensus sequence.
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Control Panning Experiment::
Approximately 2 x 1011 phage (10 µl) is diluted with 100 µl TBST and is exposed to streptavidin as a target (see The Ph.D.-12 Phage Display Peptide Library Kit Manual). To complex any biotin in the BSA, the blocking reagent is prepared by adding 0.1 µg/ml streptavidin to the standard blocking solution. The bound phage is eluted with 0.1 mM biotin in TBS for at least 30 minutes. After 3 rounds of enrichment/amplification, the consensus sequence for streptavidin-binding peptides was determined to contain the motif: H P Q (6).
Epitope mapping with the Ph.D.-12 Library::
The library was panned against anti-β-endorphin monoclonal antibody in solution, followed by affinity capture of the antibody-phage complexes onto Protein A-agarose (rounds 1 and 3) or Protein G-agarose (round 2). The results clearly show that the epitope for this antibody spans the first 4 residues of β-endorphin (YGGF), with some flexibility allowed in the third position. One clone from round 3 had no insert.
Amino Acid Distribution of the Ph.D.-12™ Library: A total of 106 clones from the naive library were sequenced. Seven clones (7%) did not contain a displayed peptide insert. The overall amino acid distribution from the sequenced codons (1188) is shown above.
References
- Parmley, S.F. and Smith, G.P. (1988) Gene, 73, 305-318.
- Smith, G.P. and Scott, J.K. (1993) R. Wu (Eds.), Methods Enzymol., 217, pp. 228-257. San Diego: Academic Press.
- Cortese et al. (1995) Curr. Opin. Biotechnol., 6, 73-80.
- Scott, J.K. and Smith, G.P. (1990) Science, 249, 386-390.
- Cwirla, S.E., Peters, E.A., Barret, R.W. and Dower, W.J. (1990) Proc. Natl. Acad. Sci. USA, 87, 6378-6382.
- Devlin, J.J., Panganiban, L.C. and Devlin, P.E. (1990) Science, 249, 404-406.
Companion Products
Ph.D.™ Peptide Display Cloning System
Ph.D.™-12 Phage Display Peptide Library Kit
Ph.D.™-7 Phage Display Peptide Library
Ph.D.™-7 Phage Display Peptide Library Kit
Ph.D.™-C7C Phage Display Peptide Library Kit
pMAL-pIII Vector
Legal
Licenses/Patents/Disclaimers:
This product is sold for research use only and not for resale in any form. Commercial use of this product may require a license. For license information under U.S. Patent 5,866,363 please contact the Licensing Office, New England Biolabs, Inc. 240 County Road, Ipswich, MA 01938.
Commercialization of sequences covered using these products may require a license from Dyax Corp. under US Patents 5,223,409, 5,403,484 and/or 5,571,698 and associated patent rights. For license information contact the Director of Corporate Development, Dyax Corp., One Kendall Square, Bldg. 600, Cambridge, MA 02139 USA. Fax 617-225-2501.
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