 | | | | Ph.D.™-C7C Phage Display Peptide Library | | | |
 |
|
This product is currently
not available for online ordering.
|
Description:
The Ph.D.™-C7C Phage Display Peptide Library is based on a combinatorial library of random heptapeptides fused to a minor coat protein (pIII) of M13 phage (1-6). Unlike other Phage Display Libraries from NEB, the randomized sequence is flanked by a pair of cysteine residues. Under nonreducing conditions the cysteines will spontaneously form a disulfide cross-link, resulting in phage display of cyclized peptides, in contrast to the linear peptides displayed in the Ph.D.-7 and Ph.D.-12 libraries. Disulfide-constrained peptide libraries (7) have proven useful in identification of structural epitopes (8,9), mirror-image ligands for D-amino acid targets (10) and leads for peptide-based therapeutics (11). The disulfide-constrained heptapeptides are expressed at the N-terminus of plll, with the first cysteine preceded by an alanine residue and the second cysteine followed by a short spacer (Gly-Gly-Gly-Ser) and then the wild-type plll sequence. The library consist of 1.2 x 109 electroporated sequences amplified once to yield approximately 200 copies of each sequence in 10 µl of the supplied phage.
Please click the link for Applications of the Ph.D. Phage Display Peptide Libraries.
Supplied in: TBS with 50% glycerol.
Complexity: 1.2 x 109 transformants.
Reaction & Storage Conditions
Storage Conditions:
50% Glycerol
Storage Temperature:
-20°C
Notes
General notes:- Supplied in: TBS with 50% glycerol.
Usage notes:- Complexity: 1.2 x 109 transformants.
FAQs
- 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.-C7C 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: C G X F/Y/W S/N H P Q C (6).
Amino Acid Distribution of the Ph.D.-C7C Library: A total of 83 clones from the native library were sequenced. All of the sequenced clones correctly contained a 7-residue peptide flanked by cysteine residues. The overall amino acid distribution from the 581 sequenced codons (83 clones x 7 randomized codons).
References
- Parmley, S.F. and Smith, G.P. (1988) Gene, 73, 305-318.
- Schumacher, T.N.M., Mayr, L.M., Minor, D.L., Milhollen, M.A., Burgess, M.W. and Kim, P.S. (1996) Science, 271, 1854-1857.
- Wrighton, N.C. et al. (1996) Science, 271, 458-463.
- 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., Barrett, 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.
- McLafferty, M.A., Kent, R.B., Ladner, R.C. and Markland, W. (1993) Gene, 128, 29-36.
- Hoess, R.H., Mack, A., Walton, H. and Reilly, T.M. (1994) J. Immunol., 153, 724-729.
- Luzzago, A., Felici, F., Tramontano, A., Pessi, A. and Cortese, R. (1993) Gene, 128, 51-57.
Companion Products
Ph.D.™ Peptide Display Cloning System
Ph.D.™-12 Phage Display Peptide Library
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.
|
