A novel, automated, and highly selective phosphopeptide enrichment for phosphopeptide identification and phosphosite localization

16 Nov 2020

Phosphopeptide enrichment is one of the most challenging sample preparation steps for LC/MS analysis due to the variation in reproducibility and selectivity caused by manual sample preparation. In this application note, Agilent Technologies performs an automated phosphopeptide enrichment with both qualitative and quantitative analysis using the Agilent AssayMAP Bravo platform and an LC/Q‑TOF system was implemented for the HUPO Phosphopeptide Challenge.

Agilent 1290 Infinity II LC System

Agilent Technologies

The 1290 Infinity II LC System is an ultra-high performance LC that has market-leading sample capacity and is designed for the lowest dispersion for the most challenging ultra-HPLC experiments. The 1290 Infinity II LC system is designed to be both a HPLC and high-end UHPLC system in a single flow path, giving you robust and excellent performance no matter what application you are running while seamlessly integrating into your existing laboratory environment.

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Protein PurificationProtein purification is a vital step in drug discovery, therapeutics, biotech and life science research. The purification process typically involves subcellular or membrane protein extraction with cell lysis kits, separation of proteins from cell debris by filtration or spin columns, and the isolation of proteins of interest from other proteins and impurities with affinity purification (including fusion protein tags and antibody binding proteins A, G and L), immunoprecipitation or chromatographic methods, such as ion exchange, size exclusion and immobilized metal affinity chromatography. All purification methods come in multiple formats for your laboratory needs, including agarose or magnetic beads, resins, columns and filter plates. Find the best protein purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.LC-MSLC-MS (liquid chromatography-mass spectrometry) systems and equipment are used for separation and quantitative analysis of complex mixtures, combining liquid chromatography and mass spectrometry. Quantify proteins, contaminants, pesticides or screen for drug metabolites with a high level of sensitivity. LC-MS systems and equipment include reverse phase, normal phase and specialized columns integrated with various MS detectors such as time-of-flight (TOF), quadrupole, orbitrap or ion trap mass analyzers. LC-MS/MS instruments equipped with a qTOF or triple quadrupole analyzer give greater sensitivity and resolving power to your analysis. Find the best LC-MS equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.AutomationAutomation in laboratories and manufacturing processes enhances efficiency, precision, and scalability by reducing the need for manual intervention. It plays a critical role in improving productivity, minimizing human error, and accelerating workflows in fields like diagnostics, drug development, and industrial testing. Automation technologies include robotic systems, automated liquid handlers, and process control systems that streamline complex tasks and ensure consistent, reproducible results. Explore our peer-reviewed product directory to discover the best automation solutions, compare options, read user reviews, and get prices directly from manufacturers.PeptidesPeptides are short chains of amino acids that play key roles in biological processes, including signaling, immune responses, and enzyme regulation. Peptide research is critical in drug discovery, diagnostics, and vaccine development. Browse our peer-reviewed product directory to find the best peptides and peptide-related products, compare options, check reviews, and get pricing directly from manufacturers.Phosphorylation