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HeavyPeptide IGNIS Prime Peptide Quantitation Kit


The next generation of standards for rapid absolute peptide quantitation.


Thermo Scientific HeavyPeptide IGNIS Prime Peptide Quantitation Kit The Thermo Scientific HeavyPeptide IGNIS Prime Peptide Quantitation Kit enables absolute quantitation of target peptides by mass spectrometry (MS) in less time than conventional methods of isotopically labeled standard-based peptide quantitation.

This kit contains a HeavyPeptide IGNIS Prime Peptide, which is a custom isotopically labeled peptide synthesized with a C-terminal, isotopically labeled Universal Reporter Peptide (URP) separated by a tryptic site (lysine or arginine); upon digestion, this dipeptide yields 2 peptides of different masses that exhibit a 1:1 stoichiometry. Each dipeptide comes with an Isotopologue Dilution Curve Mixture (iDCM-8) and Isotopologue Bracketing Mixture (iBM-4), reagents comprised of 8 and 4 isotopologues, respectively, with known concentrations that are used for single-injection instrument calibration and peptide quantitation.

Highlights:

  • Rapid – decreases time to generate concentration curve (single run vs. multiple runs by other methods)
  • Sensitive – enables the absolute quantitation of low-abundant proteins (attomolar sensitivity)
  • Precise – single-injection dilution curve removes LOD/LOQ bias
  • Flexible – multiple isotopic amino acids and reporter sizes available

Includes:

  • 10 x 1.0μg (~0.5nmol) HeavyPeptide IGNIS Prime Peptide – target peptide quantitation standard
  • 10 x Isotopologue Dilution Curve Mixture (iDCM-8) – for single-injection instrument calibration and quantitation of 1-2 target peptides
  • 10 x Isotopologue Bracketing Mixture (iBM-4) – for multiplex quantitation (up to 6 target peptides)
  • 10 x Reconstitution Solvent

Applications:

  • Selected reaction monitoring (SRM) and multiple reaction monitoring (MRM) assays
  • Biomarker discovery, verification and validation
  • Functional quantitative proteomics
  • Confirmation of results from RNA interference (RNAi)
  • Pharmacokinetics
  • Metabolomics
  • Clinical biochemistry for drug and metabolite monitoring
  • Anti-doping testing
  • Protein expression monitoring
  • Cell signal profiling and pathway validation

Product Details:

Specifications of Thermo Scientific HeavyPeptide IGNIS Prime Peptides
Dipeptide purity >95%
Isotopic purity >99%
Aliquots X Quantity 10 X 1.0μg (~0.5nmol§)
Custom peptide length •  Up to 20 amino acids in length
•  C-terminus must be arginine or lysine
•  Sequence must include a heavy amino acid
Includes: •  10 X Isotopologue Dilution Curve Mixture (iDCM-8)
•  10 X Isotopologue Bracketing Mixture (iBM-4)
•  10 X 1mL Reconstitution Solvent
As measured by analytical reverse-phase HPLC
§ The quantity provided is the gross weight of peptide, not the net peptide content, and is based on an average peptide content of 60% (corresponding to approximately 0.5 nmol).

Visit HeavyPeptide AQUA Custom Synthesis Service page for all information on peptide isotopes

Quantitative proteomics using isotope dilution mass spectrometry (IDMS) relies on isotopically labeled peptides that act as standards that mass spectrometers use to detect and quantify target proteins. HeavyPeptide IGNIS Prime peptides are dipeptides comprised of a custom HeavyPeptide sequence and a Universal Reporter Peptide (URP) separated by a tryptic residue (lysine or arginine). The URP is selected from a list of isotopologues that have an identical sequence (AALPAAFK) but vary by mass due to selective isotopic labeling.

How to build Thermo Scientific HeavyPeptide IGNIS Prime Peptides
Steps 1-3 in building Thermo Scientific HeavyPeptide IGNIS Prime Peptides
HeavyPeptide IGNIS isotopologues:
same sequence but variable mass
Isotopologue Sequence Δm§
HeavyPeptide IGNIS isotopologue iA AALPAAFK +6
HeavyPeptide IGNIS isotopologue iB AALPAAFK +11
HeavyPeptide IGNIS isotopologue iC AALPAAFK
HeavyPeptide IGNIS isotopologue iD AALPAAFK +16
HeavyPeptide IGNIS isotopologue iE AALPAAFK +21
HeavyPeptide IGNIS isotopologue iF AALPAAFK +26
HeavyPeptide IGNIS isotopologue iG AALPAAFK +31
HeavyPeptide IGNIS isotopologue iH AALPAAFK +36
HeavyPeptide IGNIS isotopologue iJ AALPAAFK +41
HeavyPeptide IGNIS isotopologue iK AALPAAFK +47
Steps 4 and 5 in building Thermo Scientific HeavyPeptide IGNIS Prime Peptides
Isotopologue availability depends on the number of peptides to be quantitated (single peptide or multiplex).
Red letters indicate isotopically labeled amino acids.
§ Compared to the iC isotopologue
 
Component details of the Thermo Scientific HeavyPeptide IGNIS Peptide Quantitation Kit
HeavyPeptide IGNIS Prime Peptide Isotopologue Dilution Curve Mixture (iDCM-8) Isotopologue Bracketing Mixture (iBM-4)
Thermo Scientific HeavyPeptide IGNIS Prime Peptide structure

Isotopologue

amol per tube

Isotopologue

amol per tube
HeavyPeptide IGNIS isotopologue iC in iDCM-8 32,805,000 HeavyPeptide IGNIS isotopologue iG in iBM-4 32,805,000
HeavyPeptide IGNIS isotopologue iD in iDCM-8 10,935,000 HeavyPeptide IGNIS isotopologue iH in iBM-4 3,645,000
HeavyPeptide IGNIS isotopologue iE in iDCM-8 3,645,000 HeavyPeptide IGNIS isotopologue iJ in iBM-4 405,000
HeavyPeptide IGNIS isotopologue iF in iDCM-8 1,215,000 HeavyPeptide IGNIS isotopologue iK in iBM-4 45,000
HeavyPeptide IGNIS isotopologue iG in iDCM-8 405,000  
Thermo Scientific HeavyPeptide IGNIS Prime Peptide structure. Custom, isotopically labeled peptide synthesized with selected isotopologue separated by a tryptic cleavage site. HeavyPeptide IGNIS isotopologue iH in iDCM-8 135,000
HeavyPeptide IGNIS isotopologue iJ in iDCM-8 45,000
HeavyPeptide IGNIS isotopologue iK in iDCM-8 15,000

Each HeavyPeptide IGNIS Prime Peptide comes with the Isotopologue Dilution Curve Mixture (iDCM-8), which is comprised of 8 isotopologues. Because each iDCM-8 aliquot has a known amount of each isotopologue, the reagent can be used to cut instrument calibration time by performing a single MS run instead of the multiple rounds of MS for each isotopologue that would normally be needed to generate the standard curve.

Single-injection instrument calibration with Thermo Scientific IGNIS Dilution Curve Mixture (iDCM-8)
Single-injection instrument calibration with Thermo Scientific Isotopologue Dilution Curve Mixture (iDCM-8).

The HeavyPeptide IGNIS Prime Quantitation Kit can be used for single-peptide or multiplex quantitation. In each case, the IGNIS Prime Peptide is combined with either iDCM-8 or the Isotopologue Bracketing Mixture (iBM-4) and spiked into the sample. During LC-MS, the iDCM-8 and iBM-4 act as internal standards to determine the quantity of the URPs and therefore the custom HeavyPeptide standard (because of the 1:1 stoichiometry).

Because each URP has a specific mass, isotopologue selection for use as the URP for a given IGNIS Prime Peptide is dependent upon the number of peptides that are to be quantitated. For single or duplex quantitation, only 2 specific isotopologues can be used because iDCM-8 is added as the internal control (which contains 8 of the 10 isotopologues). For multiplex quantitation of up to 6 peptides, a greater number of isotopologues can be used as URPs because iBM-4 only contains 4 isotopologues.

Single peptide and multiplex quantitation using the Thermo Scientific HeavyPeptide IGNIS Prime Peptide Quantitation Kit
Single-peptide and multiplex quantitation using the Thermo Scientific HeavyPeptide IGNIS Prime Peptide Quantitation Kit.

Related Resources:

Peptide Analysis Tool
Antigen Profiler for Robust Peptide Antibody Design
List of Peptide Modifications
Amino Acid Single-Letter Codes and Physical Properties

Protein Methods Library: Peptide Synthesis
Protein Methods Library: Peptide Design
Protein Methods Library: Quantitative Proteomics

Related Products:

HeavyPeptide AQUA Custom Synthesis Service
HeavyPeptide AQUA FasTrack Service
Custom Peptide Synthesis Service
1-Step Heavy Protein In Vitro Protein Expression Kits
Peptide Sample Prep Tools for Mass Spectrometry

Learn about all Peptide Products


Quote Request and Ordering Information
Download the Quote Request/Order Form (1.5mb) Download the peptide order form

Submission Information
For North America:

Email quote requests or orders to
peps@thermofisher.com

OR

Contact your local sales representative or call 1-800-874-3723 or 1-815-968-0747.

For Europe and all other regions:

Email quote requests to services.biopolymers@thermofisher.com

Email orders to sales.peptides@thermo.com

Or contact your local sales representative or Thermo Fisher Scientific in Ulm, Germany at +49-731-29870.

For technical support, please call
1-800-874-3723 or 1-815-968-0747
For technical support, please call +49-731-29870.

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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