Pierce™ Peptide Retention Time Calibration Mixture
Pierce™ Peptide Retention Time Calibration Mixture
Thermo Scientific™

Pierce™ Peptide Retention Time Calibration Mixture

The Thermo Scientific Pierce Peptide Retention Time Calibration Mixture enables reversed phase liquid chromatography (LC) and mass spectrometry (LC-MS) usersRead more
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Catalog number 88321
Price (USD)
750.00
Each
Add to cart
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Price (USD)
750.00
Each
Add to cart
The Thermo Scientific Pierce Peptide Retention Time Calibration Mixture enables reversed phase liquid chromatography (LC) and mass spectrometry (LC-MS) users to optimize and confirm correct operation of their equipment.

Features of Peptide Retention Time Calibration Mixture:

• Assess chromatography and MS instrument performance
• Predict peptide retention across multiple instrument platforms
• Predict peptide retention time from sequence using calculated hydrophobicity factor
• Optimize scheduled MS acquisition windows for improved quantification and increased multiplexing
• Normalize collision energies between instruments and platforms
• Normalize results for variation in retention times and peak intensities between runs

The Pierce Peptide Retention Time Calibration Mixture is used to optimize and assess LC parameters, identify the total peptide elution window, and optimize MS parameters. Testing the mixture frequently throughout the lifetime of a column or including it in quantitative samples makes it possible to monitor and normalize LC-MS performance between samples and over time. When used with Thermo Scientific Pinpoint 1.1 software, the calibration mixture can predict peptide retention times based upon peptide sequence or relative retention time between instruments. The mixture can also be used with Pinpoint software to establish normalized collision energies between platforms. This allows normalization of methods and eases the transfer of quantitative methods between LC-MS platforms. Pinpoint software, sample files, tutorials, and videos can be downloaded from the Thermo Proteomics Portal.

The Pierce Peptide Retention Time Calibration Mixture contains 15 synthetic heavy peptides mixed at an equimolar ratio that elute across the chromatographic gradient. The observed retention times and hydrophobicity factors (HF) for these calibrant peptides are fit to a linear equation to determine the slope of the retention time/HF relationship. This equation and the HF of an uncharacterized peptide are then used to predict retention time.

The prediction of peptide retention time assists in the development of multiplexed, high throughput mass spectrometric assays. The Pierce Peptide Retention Time Calibration Mixture and Thermo Scientific Pinpoint software streamline the transition from qualitative protein discovery results to the development of targeted MS assays on Thermo Scientific triple quadrupole, Orbitrap, Exactive, and ion trap mass spectrometers. The Pierce Peptide Retention Time Calibration Mixture permits optimization and regular assessment of chromatographic performance and for rapid development of multiplexed, scheduled targeted MS assays for the quantification of dozens to hundreds of peptide targets per run.

More Product Data
Optimize liquid chromatography with a peptide retention time calibration mixture
For Research Use Only. Not for use in diagnostic procedures.
Specifications
FormulationMixture of 15 synthetic heavy peptides at 5uM each
For Use With (Equipment)Mass Spectrometer
Product TypePeptide Retention Time Calibration Mixture
Product LinePierce™
Starting MaterialPeptides
Final Product TypePeptides
Detection MethodMass Spectrometry
Workflow StepCalibration
Quantity200 μL
FormLiquid
Unit SizeEach
Contents & Storage
Upon receipt store at -80°C.

Frequently asked questions (FAQs)

After LC-MS analysis using the Pierce Retention Time Calibration Mixture (PRTC) or Pierce LC-MS/MS System Suitability Standard (7x5 mix), the chromatography is drifting over time. How do I fix the chromatography so that it is more reproducible?

We recommend purging the LC system and checking the flow meter calibration. We also recommend checking mobile phase solvents and LC gradient. Evaluate and replace the column if needed and rerun the Pierce Retention Time Calibration Mixture (PRTC)/Pierce LC-MS/MS System Suitability Standard (7x5 mix) to verify that the chromatography is consistent in replicate runs.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.

How do I analyze the data for Pierce Retention Time Calibration Mixture (PRTC) and Pierce LC-MS/MS System Suitability Standard (7x5 mix) standard?

Create a new Skyline document containing 15 PRTC heavy isotope-labeled peptides. The Skyline document can be used for the DDA data analysis using MS1 filter or for the PRM/SRM data analysis with targeted MS filters. The Skyline document for 7x5 standard is available as a web download document. Please refer 7x5 standard label for detailed information. Alternatively, the 7x5 standard data can be analyzed in Chromeleon software using the pre-built report template (web download document).

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.

What is the difference between Pierce Retention Time Calibration Mixture (PRTC) and Pierce LC-MS/MS System Suitability Standard (7x5 mix)?

Pierce Retention Time Calibration Mixture (PRTC) is a mixture of 15 peptides with varying hydrophobicity used to determine the quality of nano and capillary flow LC chromatography and column performance. The Pierce LC-MS/MS System Suitability Standard (7x5 mix) contains 7 of 15 PRTC peptides with five isotopologue versions of each peptide containing no heavy, 1 heavy, 2 heavy, 3 heavy, and 4 heavy isotope-labeled amino acids. The isotopologues for each peptide are provided at a distinct concentration to generate seven standard curves to assess system performance. In addition to evaluation of LC chromatography, this standard is also used to assess the dynamic range and sensitivity of the LC-MS system.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.

How do I assess my LC-MS system performance?

The best way to assess the performance of an entire LC-MS system is using a mass spectrometry calibration solution, peptide retention time standards (Pierce Peptide Retention Time Calibration Mixture (PRTC), Cat. No. 88321 and Pierce LC-MS/MS System Suitability Standard (7 x 5 Mix), Cat. No. A40010), and a known standard sample, e.g., Pierce BSA Digest (Cat. No. 88341), Pierce 6 Protein Digest (Cat. No. 88342), or Pierce HeLa Protein Digest Standard (Cat. Nos. 88328, 88329) specific to your application. We recommend frequent assessment of LC-MS system performance on a routine basis to compare results over time.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.

I observe peptide retention time shifts in my liquid chromatography (LC) chromatogram. How can I check my LC performance?

We recommend using the Pierce Peptide Retention Time Calibration Mixture (Cat. No. 88321). It provides synthetic heavy peptides to diagnose and troubleshoot your LC (liquid chromatography) system and gradient.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.