How do I verify the LC-MS system after installation of a new column for proteomic applications?
Several technical replicate injections of 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) should be used to equilibrate and compare the new column to reference chromatograms.
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.
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
Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.
I'm struggling with mass spectrometry sample clean-up and feel that I'm losing peptides. Can you help me verify?
We recommend using the Pierce HeLa Protein Digest Standard (Cat. Nos. 88328, 88329) to test your sample clean-up method. Use it directly and also as a control, co-treated with the sample to check for any loss of peptides.
Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.
I have a low number of protein identifications from my protein digest sample. What can I do?
We recommend checking your mass spectrometry system performance using the Pierce HeLa Protein Digest Standard (Cat. Nos. 88328, 88329) to help determine if the problem is from the sample preparation or the LC-MS system.
Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.