Site-defined pSer, pThr, and pTyr peptides (including multi-site phosphorylation) with purification and analytics that confirm identity and purity.
Phosphorylated peptides—also called phosphopeptides or phospho-peptides—are peptides that contain one or more phosphate groups installed at defined residues, most commonly serine (pSer), threonine (pThr), or tyrosine (pTyr). A single phosphorylation adds ~+79.966 Da (HPO3) and introduces strong negative charge, which can meaningfully change conformation, binding, and assay behavior.
Bio-Synthesis provides custom phosphorylated peptide synthesis using robust SPPS workflows and route planning aligned to your goals. We routinely produce mono- and multi-phosphorylated peptides, as well as paired phospho/non-phospho controls for kinase studies, phospho-specific antibody projects, and LC–MS workflows.
Figure: Phosphopeptides model phosphorylation “on/off” states for signaling, binding, and enzyme assays. Kinase-mediated phosphorylation activates interactions, while phosphatase-driven dephosphorylation switches signaling off.
Defined residue-level phosphorylation helps avoid mixtures and supports clean interpretation.
Charge and recognition motifs can shift binding, enzyme kinetics, and chromatography.
We align purification and QC to your intended use (screening, MS standards, antibody work).
Related services: Peptide Modifications, Isotope-Labeled Peptides, Difficult Peptide Synthesis.
Protein phosphorylation is one of the most common post-translational modifications (PTMs). It is installed by kinases and removed by phosphatases, enabling rapid regulation of protein activity.
Because phosphorylation is dynamic in cells, many experiments require defined phospho-states that biology does not easily provide. Phosphopeptides let you isolate the “phosphorylated region” and test it directly.
Phosphorylation can encode “barcodes”—combinatorial patterns that change function. For complex signaling regions, multi-site phosphopeptides allow controlled experiments that compare specific phosphorylation patterns side-by-side.
Phosphorylation can turn interactions “on/off” by changing charge and conformation, rewiring binding and activity.
Defined phosphopeptides enable kinase/phosphatase assays and motif mapping without full-length proteins.
Phospho-specific antibodies and enrichment workflows perform best with validated phosphopeptide controls.
High-purity phosphorylated peptides (phosphopeptides) demand site control plus a synthesis route that protects the phosphate group, avoids side reactions, and preserves sequence integrity. We routinely support single-site and multi-site phosphorylation projects.
Use protected phospho-amino acid building blocks during SPPS for site-defined pSer/pThr/pTyr insertion.
When appropriate, phosphorylation is introduced after chain assembly to fit sequence constraints or project design.
Multi-phosphorylation can increase acidity and reduce coupling efficiency. We tune chemistry and workflow to maintain yield and purity.
We select a practical strategy based on your phosphorylation pattern (single vs multi-site), sequence behavior (aggregation/solubility), and the role of the phosphate group (binding epitope vs functional mimic). If your project requires strict isoform control, we prioritize building-block incorporation to enforce site definition.
Related: Difficult Peptide Synthesis · Peptide Modifications · Isotope-Labeled Peptides
Bio-Synthesis routinely prepares custom phosphorylated peptides (phosphopeptides) with one or multiple phosphate groups. The most common biologically relevant sites are Ser, Thr, and Tyr.
Core modification for kinase pathways and binding motifs; common in signaling and regulatory domains.
Frequently used for kinase substrate studies and functional motif mapping.
Key signaling handle for SH2/PTB interactions; used in binding assays and phospho-specific antibody validation.
We support peptides containing multiple phosphoserines, multiple phosphothreonines, multiple phosphotyrosines, or mixed combinations (e.g., pS/pT, pS/pY, pT/pY). Multi-site designs are especially useful for pathway-relevant motifs, kinase docking regions, and phosphoproteomics standards.
Expand the options below for common phosphopeptide types, typical use-cases, and design notes. Feasibility depends on sequence behavior, phosphorylation pattern, and downstream assay requirements.
single-site or multi-site
Tip: If your goal is enzyme kinetics, include flanking residues that match the natural sequence context (often improves specificity).
phospho vs non-phospho vs mutants
See also: Isotope-Labeled Peptides
non-hydrolyzable analogs, special motifs
Some programs require phosphatase-resistant or otherwise specialized analogs to stabilize a phosphorylation state. If you have a target analog or literature precedent, share it—our team will evaluate feasibility and propose a route.
This page targets: phosphorylated peptide synthesis, phosphopeptide synthesis, phosphorylated peptides, phosphopeptides, phosphoserine peptide, phosphothreonine peptide, phosphotyrosine peptide, multi-phosphorylated peptides, and custom phosphopeptide synthesis.
Substrate peptides, motif mapping, inhibitor screening, and specificity profiling.
Define dephosphorylation kinetics and substrate preferences.
Capture and quantify interactions that require phosphorylation.
Phosphopeptides are commonly used as standards and controls during method development, enrichment validation, and quantitative workflows. If your use-case is quantitation, we can evaluate isotope labels and matched light/heavy sets.
Phospho-specific antibodies must distinguish phospho from non-phospho epitopes. Defined phosphopeptides enable direct validation.
Site-defined phosphopeptides are most commonly made by incorporating protected phospho-amino acids during SPPS. In some designs, post-assembly phosphorylation may be considered—but it often requires more optimization to avoid mixtures. If you tell us your sequence and goals, we’ll recommend the most reliable approach.
If your sequence is also aggregation-prone (hydrophobic clusters, long length, repeats), consider linking to Difficult Peptide Synthesis for strategy options.
Figure: Typical workflow for phosphorylated peptide synthesis—design review through purification, QC, and documentation.
If your decisions depend on exact site assignment in multi-site constructs, tell us early—we’ll align analytics to it.
“Phosphorylated peptide” (phosphopeptide) refers to the final product you purchase and test. “Peptide phosphorylation” often refers to the biological process or enzymatic phosphorylation in assays. This page focuses on custom synthesis of defined phosphopeptides.
We routinely synthesize pSer, pThr, and pTyr at defined positions, including multiple sites within a single peptide.
Multiple phosphate groups increase acidity and can reduce coupling efficiency, complicate purification, and increase sensitivity to harsh conditions. Multi-site designs benefit from route planning and controlled coupling/deprotection parameters.
Yes—matched sets are strongly recommended for kinase, binding, or antibody studies. We can also provide mutants (S/T→A; Y→F) or phosphomimetics (S/T→D/E) when appropriate.
Typical confirmation includes analytical HPLC/UPLC and LC-MS (intact mass) when feasible. If you require explicit site confirmation (especially multi-site constructs), MS/MS can be added on request.
Many phosphopeptides are more polar and may behave differently in HPLC and solubility. We provide reconstitution guidance aligned to your sequence and intended use.
Send the peptide sequence, phosphorylation site(s) and residue type(s), quantity and purity target, and intended application. If you need matched controls or special analytics, include that as well.
Often yes (project-dependent). Common combinations include labels, handles, biotin, or isotope labels. Share your full design and constraints so we can propose a robust route.
Yes. Libraries are common for kinase specificity profiling, motif scanning, and antibody epitope mapping. We can recommend panel design and consistent QC for comparability.
Purity targets are set by your use-case (screening vs structural vs MS standards). We’ll recommend a realistic specification and purification plan aligned to your application.
Phosphorylated peptides (phosphopeptides) are often used as assay substrates, binding motifs, or validated controls. The table below provides practical starting points. If your workflow is sensitive to batch-to-batch variation, ask for matched non-phosphorylated controls and sequence variants.
Not sure what to order? Tell us your downstream method (assay, antibody, LC–MS), and we’ll recommend a practical set (phospho + non-phospho + variants) and a QC plan aligned to it.
If you have a kinase motif, substrate region, or UniProt residue numbering, include it—we can help translate that into an order-ready sequence.
Use this checklist to avoid “looks fine on paper” failures in assays.
Bio-Synthesis is optimized for custom phosphorylated peptide synthesis (phosphopeptides) with defined sites, fit-for-purpose purification, and documentation that supports reproducibility.
For the fastest quote, send your sequence(s), phosphorylation site(s) (pSer/pThr/pTyr), desired quantity and purity, and intended use (kinase assay, binding, antibody, LC–MS). We’ll recommend a practical synthesis/purification plan plus fit-for-purpose QC.
What happens next: Our technical team reviews your request and responds with feasibility notes, recommended route options, a QC plan, and pricing.
Not sure what to request? Send the biological context (kinase/phosphatase target or pathway) and we’ll recommend a practical peptide design and QC plan.
Selected peer-reviewed references on phosphorylation “barcodes,” multi-site phosphopeptide synthesis challenges, and practical strategies.
References are provided for background and design insight; Bio-Synthesis does not claim ownership of the cited works.
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