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Biochemistry

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JoVE Journal Biochemistry
Sample Preparation and Relative Quantitation using Reductive Methylation of Amines for Peptidomics Studies
 

Sample Preparation and Relative Quantitation using Reductive Methylation of Amines for Peptidomics Studies

Article DOI: 10.3791/62971-v • 08:00 min • November 4th, 2021
November 4th, 2021 •
1,2, 1, 1, 1,2
1Department of Biological and Environmental Sciences, Bioscience Institute, Sao Paulo State University (UNESP), 2Biodiversity of Coastal Environments Postgraduate Program, Bioscience Institute, Sao Paulo State University (UNESP)

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This article describes a sample preparation method based on heat-inactivation to preserve endogenous peptides avoiding degradation post-mortem, followed by relative quantitation using isotopic labeling plus LC-MS.

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Sample Preparation Relative Quantitation Reductive Methylation Amines Peptidomics Studies Fast Heat Inactivation Protease Intracellular Peptides Degradation Isotopic Labeling Commercially Available Reagents Inexpensive Chemically Stable Serum Samples Human Neuroblastoma Cells DMEM FBS Penicillin Streptomycin PBS Centrifugation Supernatant Pellet Ultra Purified Deionized Water Zebrafish Brain Freeze At Minus 80 Degrees Celsius Tissue Sample Resuspension Sonication
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