Stable Isotope-Labeled (SIL) Peptides & Heavy Isotope Labeled Peptide Standards

Defined heavy labels (13C/15N/D) and TMT6 options for quantitative LC-MS/MS, method validation, and structural analysis workflows.

Build accurate internal standards. Match digestion and chromatography, separate light vs heavy by mass, and report confidently.

Overview: Stable Isotope-Labeled (SIL) Peptides

What are SIL peptides?

Stable isotope-labeled (SIL) peptides—also referred to as isotope-labeled peptides or heavy isotope-labeled peptides—incorporate ^13C, ^15N, and/or deuterium (D) at defined amino acid residues to introduce a predictable mass shift (Δm) without altering the peptide sequence. In LC-MS/MS workflows, the labeled (heavy) peptide closely matches the chromatographic behavior of the native (light) peptide while remaining distinguishable by mass, enabling accurate and reproducible quantitation.

Bio-Synthesis synthesizes SIL peptides as matched light/heavy pairs using ^13C/^15N or deuterium labeling, with optional TMT6 channels for multiplexed analysis. Peptides are delivered LC-MS/MS-ready with purification, analytical QC, and certificates of analysis (COA) to support absolute quantitation and structural analysis workflows.

Stable isotope-labeled peptide standards supplied by Bio-Synthesis, Inc. (Lewisville, TX) have been reported in peer-reviewed Molecular & Cellular Proteomics literature for targeted quantitative proteomics applications.

ISO 900:2015/ISO13485:2016 45+ Years of Expertise U.S. Facilities - Texas 13C/15N & D labeling TMT6 N-term tags HPLC/LC-MS QC
Why choose Bio-Synthesis
  • Design-first quoting: we align label choice to digestion (tryptic Lys/Arg), enrichment, and method goals.
  • Site-defined labeling: internal heavy residues or N-terminal tagging (e.g., TMT6-126 to TMT6-131).
  • Fit-for-purpose QC: analytical HPLC/UPLC + LC-MS where feasible; COA that supports method validation.
  • Peptide expertise: difficult, hydrophobic, and modified sequences handled with practical synthesis strategy selection.
Light vs heavy: what changes

Same sequence and chemistry; a controlled mass shift (Δm) enables absolute or relative quantitation.

  • Light peptide: unlabeled reference (endogenous or synthetic)
  • Heavy peptide: SIL standard (e.g., [K(13C6;15N2)], [R(13C6;15N4)])
  • Quant readout: peak area ratio (heavy/light) after matched processing
Color schematic of SIL peptide standard workflow showing matched light/heavy peptides and LC-MS/MS quantitation

Figure: Color SIL peptide standard workflow—matched light/heavy peptides with a defined mass shift (Δm) enable robust LC‑MS/MS quantitation.

Related: Isotope-Labeled Peptides, Peptide Modifications, Click Chemistry Peptides. If your sequence is challenging, see difficult peptide synthesis. For quick turnaround, browse ready-made catalog peptides.

Isotopic enrichment & label options (keyword table)

Use this table to copy/paste the exact label keyword you want in your quote request. Mass differences are the nominal heavy–light shifts for the labeled residue or tag. Final peptide mass and labeling confirmation are reported on the COA.

Type Name Abbreviation / alternative name Mass difference (ΔDa) Isotopic enrichment Position
Isotopic Heavy Ala (13C3;15N) [A(13C3; 15N)] +4.007 ≥98 atom% (typical) Internal
Isotopic Heavy Ala (D3) [A(D3)] +3.019 ≥98 atom% (typical) Internal
Isotopic Heavy Arg (13C6) [R(13C6)] +6.020 ≥98 atom% (typical) Internal
Isotopic Heavy Arg (13C6;15N4) [R(13C6; 15N4)] +10.008 ≥98 atom% (typical) Internal
Isotopic Heavy Asn (13C4;15N2) [N(13C4; 15N2)] +6.008 ≥98 atom% (typical) Internal
Isotopic Heavy Asp (13C4;15N) [D(13C4; 15N)] +5.011 ≥98 atom% (typical) Internal
Isotopic Heavy Gln (13C5;15N2) [Q(13C5;15N2)] +7.011 ≥98 atom% (typical) Internal
Isotopic Heavy Glu (13C5;15N) [E(13C5; 15N)] +6.014 ≥98 atom% (typical) Internal
Isotopic Heavy Gly (13C2;15N) [G(13C2; 15N)] +3.004 ≥98 atom% (typical) Internal
Isotopic Heavy His (15N3) [His(15N3)] +2.991 ≥98 atom% (typical) Internal
Isotopic Heavy Ile (13C6;15N) [I(13C6; 15N)] +7.017 ≥98 atom% (typical) Internal
Isotopic Heavy Leu (13C1) [L(13C1)] +1.003 ≥98 atom% (typical) Internal
Isotopic Heavy Leu (13C6) [L(13C6)] +6.020 ≥98 atom% (typical) Internal
Isotopic Heavy Leu (13C6;15N) [L(13C6; 15N)] +7.017 ≥98 atom% (typical) Internal
Isotopic Heavy Leu (D10) [L(D10)] +10.063 ≥98 atom% (typical) Internal
Isotopic Heavy Leu (D3) [L(D3)] +3.019 ≥98 atom% (typical) Internal
Isotopic Heavy Lys (13C6) [K(13C6)] +6.020 ≥98 atom% (typical) Internal
Isotopic Heavy Lys (13C6;15N2) [K(13C6; 15N2)] +8.014 ≥98 atom% (typical) Internal
Isotopic Heavy Phe (13C9;15N) [F(13C9; 15N)] +10.027 ≥98 atom% (typical) Internal
Isotopic Heavy Pro (13C5;15N) [P(13C5; 15N)] +6.014 ≥98 atom% (typical) Internal
Isotopic Heavy Ser (13C3;15N) [S(13C3;15N)] +4.007 ≥98 atom% (typical) Internal
Isotopic Heavy Ser (D3) [S(D3)] +3.019 ≥98 atom% (typical) Internal
Isotopic Heavy Thr (13C4;15N) [T(13C4;15N)] +5.011 ≥98 atom% (typical) Internal
Isotopic Heavy Trp (13C11;15N2) [W(13C11;15N2)] +13.031 ≥98 atom% (typical) Internal
Isotopic Heavy Tyr (13C9;15N) [Y(13C9; 15N)] +10.027 ≥98 atom% (typical) Internal
Isotopic Heavy Val (13C5) [V(13C5)] +5.017 ≥98 atom% (typical) Internal
Isotopic Heavy Val (13C5;15N) [V(13C5; 15N)] +6.014 ≥98 atom% (typical) Internal
Isotopic L-tyrosine-phenyl-d4 (deuterated Tyr) [Tyr(D4)] +4.025 ≥98 atom% (typical) Internal
Isotopic TMT6-126 TMT6-126-NH +229.163 (nominal) Reagent-defined N-term
Isotopic TMT6-127 TMT6-127-NH +229.163 (nominal) Reagent-defined N-term
Isotopic TMT6-128 TMT6-128-NH +229.163 (nominal) Reagent-defined N-term
Isotopic TMT6-129 TMT6-129-NH +229.163 (nominal) Reagent-defined N-term
Isotopic TMT6-130 TMT6-130-NH +229.163 (nominal) Reagent-defined N-term
Isotopic TMT6-131 TMT6-131-NH +229.163 (nominal) Reagent-defined N-term

Tip: if your assay uses trypsin, heavy Lys/Arg are often the default internal-standard choices. For multiplexed workflows, specify the desired TMT6 channel in your quote request.

Stable Isotope-Labeled Peptides in Structural Analysis

MS-based structural workflows

SIL peptides are commonly used to turn a structural readout into a quantitative readout.

  • Crosslinking-MS & footprinting: quantify site occupancy or compare conditions with heavy standards.
  • PTM mapping: heavy modified peptides calibrate phosphorylation/acetylation/glycosylation site abundance.
  • Conformation or interaction studies: targeted PRM/MRM on signature peptides supports comparative structural claims.
NMR and isotope-edited experiments (project-dependent)

When your goal is resonance assignment or simplified spectra, heavy labeling can be useful.

  • 13C/15N labeling supports isotope-edited NMR approaches for peptides and peptide–ligand interaction studies.
  • Label placement matters: choose residues that directly report on the region of interest.
  • Confirm on COA: enrichment and identity verification are documented for downstream reporting.

Tell us the technique (NMR, MS, HDX, crosslinking) and the acceptance criteria; we’ll align the label plan to it.

Applications of Stable Isotope-Labeled Peptides

AQUA & absolute quantitation

Heavy peptide spikes provide known amounts to convert MS signal into concentration.

  • Biomarker validation
  • PK/PD readouts
  • Method transfer/verification
Targeted LC-MS/MS

MRM/SRM and PRM assays rely on matched heavy internal standards.

  • Transition optimization
  • Matrix effects control
  • Batch-to-batch comparability
DIA benchmarking & libraries

Use SIL peptides to validate identification and quantify performance across runs.

  • Retention time anchors
  • Instrument performance tracking
  • Multi-lab comparability
TMT workflows (TMT6-126 to TMT6-131)

Tandem mass tags enable multiplexed relative quantitation; specify channel in your request.

  • N-terminal tagging (project-dependent)
  • Compatible with most proteomics workflows
  • Channel selection: 126/127/128/129/130/131
Assay controls & reference materials

SIL peptides are used as system suitability checks and internal QC controls.

  • Spike-in controls
  • Stability studies
  • Quantitation across timepoints

Modifications (combine SIL with functional chemistry)

Stable isotope labeling is frequently combined with chemical modifications to create application-ready standards. Share your target workflow (enrichment, digestion, immuno-capture, click, etc.) and we’ll propose a synthesis and QC plan.

PTM standards
  • Phosphorylation (Ser/Thr/Tyr)
  • Acetylation / methylation
  • Amidation, pyroglutamate
Handles & capture
  • Biotin
  • Azide / alkyne (click handles)
  • Fluorescent tags (project-dependent)
Structure control
  • Disulfide or thioether cyclization
  • Stapling (project-dependent)
  • PEG/spacer tuning for solubility

See also: Peptide Modifications and Click Chemistry Peptides.

Quality control & typical deliverables

Standard QC
  • Analytical HPLC/UPLC purity profile
  • Identity confirmation (LC-MS when feasible)
  • Certificate of Analysis (COA)
Label confirmation
  • Expected mass shift (Δm) reporting
  • Isotopic enrichment documented as specified
  • Sequence + modification verification
Optional add-ons

Project-dependent add-ons can be aligned to your assay sensitivity and regulatory context.

  • Custom formulation / aliquoting
  • Stability/handling recommendations
  • Method development support (scope-dependent)

FAQ

Are stable isotope-labeled peptides used as internal standards?

Yes. SIL peptides are commonly used as internal standards because the heavy peptide closely matches the native (light) peptide’s chemistry and retention while differing by a defined mass shift (Δm), enabling robust LC–MS/MS quantitation.

Which label should I choose for tryptic peptides?

For tryptic peptides, heavy Lys and/or Arg labels are commonly used so the heavy residue appears at the C-terminus after digestion. If you share your protease and peptide list, we’ll recommend the most robust label plan.

Can you supply both light and heavy peptides as a matched pair?

Yes. Matched light/heavy pairs are frequently used for assay development, standard curves, and inter-run normalization.

Does deuterium (D) labeling change retention time?

It can, depending on sequence and LC conditions. If retention matching is critical, 13C/15N labels are often preferred.

Do you support TMT6 labeling?

Yes. We can provide peptides with TMT6-126 through TMT6-131 N-terminal tags (project-dependent). Specify channel and any additional modifications.

What purity should I request?

For quantitative standards, higher purity reduces interference. Many targeted MS assays use ≥95% purity; some applications accept lower purity with appropriate characterization. Tell us your detection limits and matrix and we’ll recommend a practical target.

Can you combine SIL with PTMs (e.g., phosphopeptides)?

Yes. SIL phosphopeptides and other modified standards are supported (project-dependent). Include the exact modification site(s) and any enrichment step (IMAC/TiO2, antibodies) so we can align synthesis and QC.

Contact & quote request

For the fastest quote, send your peptide sequence(s) and any modifications, your preferred label keyword (copy from the enrichment table above), quantity (mg) and purity target, intended application (AQUA/MRM/PRM/DIA/TMT/structural analysis), and any required enrichment specification. We’ll respond with feasibility notes, a recommended synthesis/QC plan, and pricing.

Fastest path

What happens next: Our technical team reviews your request and replies with recommended labeling strategy, purification/QC plan, and delivery format aligned to your assay.

Fast quote checklist
  • Peptide sequence(s) + any PTMs and terminal state (free vs capped)
  • Desired heavy label (e.g., [K(13C6; 15N2)]) or TMT6 channel
  • Quantity (mg) + purity target
  • Intended use: AQUA / MRM / PRM / DIA / TMT / structural analysis
  • Any enrichment spec (e.g., ≥99 atom%) or special documentation needs

Recommended reading

References for stable isotope-labeled peptides, absolute quantitation, and multiplexed tagging in proteomics.

  • Quantitative proteomics using stable isotope–labeled peptide internal standards
    Molecular & Cellular Proteomics (2020). This peer-reviewed study reports the use of stable isotope–labeled peptide internal standards synthesized by Bio-Synthesis, Inc. (Lewisville, TX) for targeted LC–MS/MS–based quantitative proteomics (ScienceDirect ID: S1535947620335064).
  • Absolute quantification of proteins and phosphoproteins from cell lysates by tandem MS
    Gerber SA, Rush J, Stemman O, Kirschner MW, Gygi SP. PNAS (2003). Introduction of the AQUA strategy using synthetic stable isotope–labeled peptides for absolute quantitation.
  • Stable isotope labeling by amino acids in cell culture (SILAC)
    Ong S-E, Blagoev B, Kratchmarova I, et al. Molecular & Cellular Proteomics (2002). Foundational work establishing stable isotope labeling for quantitative proteomics.
  • Isobaric tagging for relative and absolute quantitation (iTRAQ/TMT concepts)
    Ross PL, Huang YN, Marchese JN, et al. Molecular & Cellular Proteomics (2004). Development of isobaric tagging strategies for multiplexed proteomic quantitation.
  • Tandem mass tags: a novel quantification strategy for comparative analysis
    Thompson A, Schäfer J, Kuhn K, et al. Analytical Chemistry (2003). Description of TMT labeling chemistry and multiplexed quantitative MS workflows.

References are provided for background. Bio-Synthesis does not claim ownership of the cited works.

Why Choose Bio-Synthesis

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