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KEAP1 mass spectrometry analysis For mass spectrometry analysis, 10 g of recombinant human KEAP1 (#11981-H29B-100, Sino Biologicals, Beijin, PR China) were diluted to 100 l with 20 mM Tris pH 8.0 and diafiltrated by means of a Amicon Ultra-0.5 ml device (Sigma-Aldrich, Milan, Italy) to eliminate GSH, and then treated with 30 mM M DTT and incubated 30 min at room temperature to reduce the disulfide bridges, diafiltrated to remove the excess of DTT and diluted in 20 mM Tris pH 8.0

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A binary solvent system was used, in which mobile phase A consisted of 2 mM Triisobutylamine adjusted with 5 mM acetic acid in 100% water, and mobile phase B of 100% methanol

Frequently Asked Questions: Anti-Aging Peptide Stack Research Q: What makes Epithalon unique in anti-aging peptide research

doi: 10.3390/cancers13133225 103 BranzkNGronkeKDiefenbachA

Sample preparation for LC-MS/MS typically involves steps such as protein precipitation or liquid-liquid extraction to isolate the target vitamins from the complex biological matrix before chromatographic separation and subsequent mass spectrometric detection (Konieczna et al., 2016