SPOTs technology enables researchers to generate the enormous sequence diversity of custom peptide libraries for mapping applications. This technology enables easy identification of the functional domains of proteins by conducting functional assays on peptide libraries attached to cellulose membranes.
SPOTs technology was originally developed as a system for easily determining the amino acid sequence of a peptide antigen. Peptide sequences corresponding to the specificity of an antibody are identified by conventional enzyme-linked or autoradiographic detection methods. SPOTs technology has since progressed to include a variety of new applications for research in the areas of immunology, pharmacology, enzymology, molecular biology, peptide chemistry and cell biology, which include:
- Combinatorial peptide library
- Protein-protein and receptor-ligand interactions
- Phosphorylation studies
- Metal binding studies
- Nucleic acid binding
- Allergen determination
Custom SPOTs Service
Sigma-Genosys also offers a custom SPOTs service. We offer formats for any number of custom peptides to be synthesized on cellulose membranes using the same chemistry as in our SPOTs kit.
Peptide synthesis is performed by automated instrumentation and individual coupling reactions are followed by monitoring the fluorescence absorption of the Fmoc protecting group as well as colorimetric staining of the free amine on the spots. After the final cycle, peptides are N-terminally aceytlated and then side chain deprotected. The fidelity of the peptide synthesis is monitored by synthesis of a standard control peptide that is probed with a known antibody. A duplicate control peptide is synthesized and cleaved from the membrane and analyzed by mass spectroscopy and HPLC for purity.
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