Hplc Peptide Standards

Peptide Retention Standard Thermo Scientific™

Thermo Scientific Pierce Peptide Retention Standard is a mixture of five c-terminal amide decapeptides that provide reference elution profiles in reverse-phase HPLC.

Features of Peptide Retention Standard:

• Determine the relative order of peptide elution from a complex mixture
• Efficiently predict peptide elution profiles to simplify identification and save purification time
• Identify peptides of known amino acid composition on reverse phase HPLC columns
• Monitor column performance—efficiency, selectivity and resolution during column aging
• Evaluate reverse-phase supports of varying n-alkyl chain lengths and ligand densities
• Compare reverse-phase columns from different manufacturers

Peptide Retention Standard allows accurate prediction of elution time for peptides of known amino acid composition up to 20 residues in length. The reference standard helps to predict the relative order of peptide elution within a complex mixture, simplifying identification of specified peptides and saving time in peptide purification. By testing the standard frequently throughout the lifetime of a column, it is possible to monitor performance characteristics such as efficiency, selectivity, and resolution during column aging. The standard can also be used to evaluate different reverse phase supports of varying n-alkyl chain lengths and ligand density, as well as columns from different manufacturers.

The Peptide Retention Standard consists of five C-terminal amide decapeptides, denoted S1 to S5, having the generic formula Ac-Arg-Gly-X-X-Gly-Leu-Gly-Leu-Gly-Lys-Amide. Four of these peptides are Na-acetylated with the sequence variation as follows: Gly3-Gly4, Ala3-Gly4, Val3-Gly4 and Val3-Val4. The fifth peptide, Ala3-Gly4, contains a free Na-amino group. This mixture will provide 100-200 injections (approximately 0.1 mg of each peptide) at 0.1 AUFS at 210nm. The Peptide Retention Standard is supplied as a dry film in a sealed vial.

A simple, quantitative method for predicting peptide retention times was developed by Guo, et al. (1985, 1986). Retention times are predicted by totaling the values that represent the contribution in minutes of each amino acid residue and the peptide terminal groups.

Retention time is dependent upon the molecular weight of the peptide. The effect on retention is relatively unimportant with a small peptide, but it increases with the size of the molecule. The accuracy of predicting peptide retention time significantly decreases beyond 20 residues. To ensure accuracy, a peptide standard is used to correct for instrument variation, column aging, n-alkyl chain length variation and ligand density.

Pierce™ Peptide Retention Time Calibration Mixture Thermo Scientific™

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

Pierce™ Peptide Retention Time Calibration Mixture Thermo Scientific™

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
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