Purpose: Evaluate the reproducibility of serial protein extraction from one sample on an individual or multiple protein extraction.
Methods: Perform HR/AM MS analysis of MSIA D.A.R.T serial extractions from a single sample using one Ab for a single protein or multiple Abs (one per D.A.R.T tip).1 The relative quantitation of peptides was used to determine depletion and disruption effects from serial dilution.
Results: Pooling of serial extractions increased targeted peptide response ca. 5-10x without significantly increasing background interference. The increased abundance resulting from pooling extractions facilitated greater protein sequence coverage and detection of N- and O-linked glycopeptides and corresponding glycoforms.
BRIMS
United States
