Retention time standard for HPLC instrument calibration
RePLiCal comprises of unique, easy to detect peptides with selected chemical properties.
- High number of calibrant points, covering a broad range of RTs
- Stable RTs over long analysis times, allowing the prediction of RTs for other peptides in a timely manner
- Includes two highly hydrophilic peptides for testing of column trapping performance
RePLiCals peptides have a high degree of linearity and thus can be used to predict the retention times of the peptides on a new gradient. By calculating iRTs, retention time windows can be adjusted for different gradients.
This tutorial demonstrates how RePLiCal can be used to re-calculate retention times using Skyline.
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RePLiCal is a HPLC calibration standard that is buildt from 27 lysine-terminating calibrant peptides. The peptides do not contain methionine, tryptophan, cysteine, and N-terminal glutamine residues. Also aspartic acid-proline, asparagine-proline and asparagine-glycine motifs are absent. It can be applied to calibration of LC gradients and nano uHPLC instruments using trap-elute configurations, real-time scheduling of selected reaction monitoring (SRM) experiments and recalibration of the m/z scale of mass spectrometry (MS) data post-acquisition. Find more information in the original publication.
1 µg RePLiCal or 25510 fmol (digested or undigested) equals 250 – 500 injections (50 – 100 fmol/injection).
Watch our tutorial to learn “How to use the RePLiCal RT standard to adapt for RT shifts in Skyline”.
- RePLiCal: A QconCAT Protein for Retention Time Standardization in Proteomics Studies. Holman S. et al., J Proteome Res. 2016 Mar 4;15(3):1090-102. [PubMed]
- Platform Independent Protein-Based Cell-Of-Origin Subtyping of Diffuse Large B-cell Lymphoma in Formalin-Fixed Paraffin-Embedded Tissue. Reinders J., et al., Sci Rep. 2020 May 12;10(1):7876. [PubMed]