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Substrate Peptides

Synthetic peptide substrates for protease, peptidase, and kinase assays — fluorogenic and chromogenic designs for measuring enzyme activity and screening inhibitors.
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Fluorogenic & Chromogenic Reporters

Built with the right reporter for your readout: AMC and AFC for fluorescence, FRET donor-quencher pairs for continuous protease assays, and pNA for chromogenic detection.



Validated for Assay Use

Each substrate is purified by HPLC and confirmed by mass spectrometry, and ships with a COA, HPLC chromatogram, and MS report, so it performs consistently in kinetic and screening work.



Custom Substrate Design

Tell us the enzyme and the cleavage site, and we design and synthesize a custom substrate, including the reporter, the spacer, and any modification you need.

FAQs

What are peptide substrates used for?

Peptide substrates are synthetic peptides cleaved or modified by a specific enzyme, used to measure that enzyme's activity. Researchers use them to run kinetics, screen inhibitors, set up high-throughput assays, and build diagnostic tests.

What is the difference between fluorogenic and chromogenic substrates?

Both report enzyme activity, through different signals. Fluorogenic substrates release a fluorophore, such as AMC, when cleaved, giving a sensitive fluorescent readout. Chromogenic substrates release a colored group, such as pNA, that is read by absorbance. Fluorogenic assays are generally more sensitive; chromogenic ones need only a plate reader.

Which enzymes can you make substrates for?

We make substrates for the main enzyme classes: serine, cysteine, aspartic, and metalloproteases, exopeptidases such as DPP-IV and aminopeptidases, and kinases. Examples include caspase, cathepsin, trypsin, chymotrypsin, and MMP substrates.

Can you design a custom substrate for my enzyme?

Yes. Send the enzyme and its cleavage specificity, and we design the substrate around the scissile bond, choose the reporter and spacer, and synthesize it to your purity. We can also work from a published sequence.

What documentation comes with each substrate?

Each substrate ships with a COA, an HPLC chromatogram, and an MS report. For assay work we can add details such as the reporter, the quencher pair, and solubility guidance on request.

What reporter groups do you offer?

For fluorescence, AMC, AFC, and FRET donor-quencher pairs such as EDANS and DABCYL. For chromogenic detection, pNA. We also add biotin or other tags when an assay needs capture or immobilization.

Understanding Peptide Substrates

A peptide substrate is a short synthetic peptide that a specific enzyme recognizes and cleaves or modifies. By attaching a reporter group that changes signal when the enzyme acts, a substrate turns enzyme activity into something you can measure: a rise in fluorescence, a color change, or a shift you can read on a plate. Peptide substrates are core reagents in enzymology, drug screening, and diagnostics.

What a Peptidase Acts On

Peptidases, also called proteases, are enzymes that cleave the peptide bonds between amino acids. Their natural substrates are peptides and proteins, and each peptidase prefers a particular sequence around the bond it cuts. A synthetic peptide substrate copies that preferred sequence and adds a reporter, so the enzyme cleaves the substrate the way it would cleave its natural target, but with a readable signal.

Fluorogenic and Chromogenic Substrates

The reporter defines the assay. Fluorogenic substrates carry a fluorophore, often AMC or AFC, that is held dark until the enzyme cleaves it free, giving a sensitive fluorescent signal. FRET substrates instead place a fluorophore and a quencher on either side of the cleavage site, such as an EDANS and DABCYL pair, so fluorescence rises as the peptide is cut, which suits continuous protease assays. Chromogenic substrates release a colored group such as pNA and are read by absorbance, which needs only a standard plate reader.

Substrates by Enzyme Class

Substrate design follows the enzyme:

  • Serine proteases, such as trypsin, chymotrypsin, and thrombin, cleave after specific residues and pair well with AMC or pNA reporters.
  • Cysteine proteases, such as caspases and cathepsins, are studied with sequence-specific fluorogenic substrates.
  • Aspartic and metalloproteases, such as BACE1 and the MMPs, are usually run with FRET substrates.
  • Exopeptidases, such as DPP-IV and aminopeptidases, use short substrates that report removal of terminal residues.
  • Kinases use peptide substrates that carry a phospho-acceptor site for activity and inhibitor assays.

Designing a Custom Substrate

A good substrate starts from the enzyme's cleavage specificity, the residues it reads on each side of the bond it cuts. We build the recognition sequence, place the reporter so it gives a clean signal on cleavage, add a spacer where needed, and synthesize to the purity your assay requires. Tell us the enzyme and the readout, and we will design and make it. For the underlying chemistry, see custom peptide synthesis and peptide modification.

Substrate peptides are supplied for laboratory research use.

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Online Consultation Email: dora@synpeptide.com Tel: +86 135 0517 2290 WhatsApp: +86 135 0517 2290