BioTides™
Biotinylated peptides are used in a variety of biomedical screening systems requiring immobilization of at least one of the interaction partners onto streptavidine coated beads, membranes, glass slides or microtiter plates. By means of its proprietary SPOT synthesis technology, JPT Peptide Technologies provides access to large numbers of biotinylated peptides at unmatched prices and fastest delivery time.
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Applications
- Identification and optimization of kinase-, phosphatase-, acetyltransferase- and histone deacetylase-substrates via standard screening systems (AlphaScreen, FlashPlates, SPA-Beads, and many more
- Mapping of protein/protein interaction sites (ELISA-like assays, precipitation of interacting proteins).
- Production of peptide microarrays.
- Loading of columns for affinity chromatography.
Example: Human c-Jun derived BioTides and protein tyrosine kinase c-Abl
Used reagents and materials:
- 384 well microtiter plate containing 160 different BioTides derived from human c-Jun protein
- Abl-kinase (New England BioLabs, www.neb.com, item# P6050L)
- Kinase buffer 50 mM Tris-HCl, 10 µM MgCl2 1 mM EGTA, 2 mM DTT, and 0.01% Brij 35 (pH 7.5, 25°C)
- SAM-Biotin capture membrane (Promega, # V7861)
- 384 pin replicator (Genetix, 384-pin QReps-PP, Product Code: X5050, printing volume approx: 0.2 µl)
| pS/pT/pY positive controls | * pS=phospho-Ser; pT=phospho-Thr; pY=phospho-Tyr |
| negative controls | |
| G8-H1 | known phosphorylation-site: Barila D, Mangano R, Gonfloni S, Kretzschmar J, Moro M, Bohmann D, Superti-Furga G.; A nuclear tyrosine phosphorylation circuit: c-Jun as an activator and substrate of c-Abl and JNK; EMBO J. 2000 Jan 17;19(2):273-81 |
Selected References
Extensive phosphorylation with overlapping specificity by Mycobacterium tuberculosis serine/threonine protein kinases (abstract)
Prisic et al., PNAS (2010)
More references under Publications/Literature
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