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Thermo Scientific™ Pierce™ Peptide Retention Time Calibration Mixture

Catalog No. PI88321
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PI88321 200 μL
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Catalog No. PI88321 Supplier Thermo Scientific™ Supplier No. 88321
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Description

15 synthetic heavy peptides mixed in equimolar ratio to test and calibrate reverse-phase liquid chromatography (LC) and mass spectrometry (LC-MS) equipment.

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.

The Pierce Peptide Retention Time Calibration Mixture is used to optimize and assess LC parameters, identify 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 is used to predict peptide retention times based upon peptide sequence and ease the transfer of methods between LC-MS platforms.

Highlights:

  • Assessment of chromatography and MS instrument performance
  • Prediction of peptide retention across multiple instrument platforms
  • Prediction of peptide retention time from sequence using calculated hydrophobicity factor
  • Optimization of scheduled MS acquisition windows for improved quantification and increased multiplexing
  • Internal standard to normalize for variation in retention times and peak intensities between runs

Specifications

Formulation Mixture of 15 synthetic heavy peptides at 5uM each
For Use With (Equipment) Mass Spectrometer
Product Type Peptide Retention Time Calibration Mixture
Product Line Pierce
Content And Storage Upon receipt store at -80°C.
Starting Material Peptides
Final Product Type Peptides
Detection Method Mass Spectrometry
Workflow Step Calibration
Quantity 200 μL
Form Liquid
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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.
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).
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.

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.
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.
Do you offer a product that will help me determine if my liquid chromatography (LC) is working correctly for peptide samples?

Yes, the Pierce Peptide Retention Time Calibration Mixture (Cat. No. 88321) can be used. It provides synthetic heavy peptides to diagnose and troubleshoot your LC (liquid chromatography) system and gradient.
Which standards and calibrants do you recommend using for assessing LC-MS system suitability?

Here are our recommendations:

- Pierce Peptide Retention Time Calibration Mixture (Cat. No. 88321) for LC system and gradient optimization
- Pierce BSA Protein Digest, MS grade (Cat. No. 88341), Pierce 6 Protein Digest, equimolar, LC-MS grade (Cat. No. 88342), and Pierce HeLa Protein Digest Standard (Cat. Nos. 88328, 88329) for "bottom up" LC-MS method optimization and LC-MS suitability assessment
- Pierce LC-MS/MS System Suitability Standard (7 x 5 Mix) (Cat. No. A40010) for assessing the gradient, sensitivity, and dynamic range of the LC-MS system for discovery and targeted peptide quantitative workflows
- Pierce TMT11plex Yeast Digest Standard (Cat. Nos. A40938, A40939) for assessing system performance for TMT-based quantitative workflows
- Pierce Intact Protein Standard Mix (Cat. Nos. A33526, A33527) for "top down" LC-MS method optimization and LC-MS suitability assessment

For Research Use Only. Not for use in diagnostic procedures.