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Tandem Mass Spectrometry01:21

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Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...
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Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
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Mass spectrometry is a powerful characterization technique that can identify and separate a wide variety of compounds ranging from chemical to biological entities, based on their mass-to-charge ratio (m/z). The instruments that allow this detection, known as mass spectrometers, have three components: an ion source, a mass analyzer, and a detector. These spectrometers differ based on the nature of their ion source and analyzers.Matrix-assisted laser desorption ionization (MALDI) is a commonly...
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Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
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Mass spectrometry is an analytical technique used to determine the molecular mass and molecular formula of a compound. The basic principle of mass spectrometry is to generate ions from the analyte molecule and measure these ion abundances against their molecular mass. One common type of ionization, known as electron ionization or EI, bombards the analyte molecules in the gas phase with high-energy electron beams. The electron beams displace an electron from the molecule and leave behind a...
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This lesson details the instrumentation of a mass spectrometer—a physical instrument to perform mass spectrometry on analyte molecules and record the characteristic mass spectra. This is achieved via three chief functions:
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Mass Spectrometry-Based Biomarker Discovery.

Weidong Zhou1, Emanuel F Petricoin2, Caterina Longo2,3

  • 1Center for Applied Proteomics and Molecular Medicine, George Mason University, 10920 George Mason Circle, MS1A9, Manassas, VA, 20110, USA. wzhou@gmu.edu.

Methods in Molecular Biology (Clifton, N.J.)
|May 15, 2017
PubMed
Summary
This summary is machine-generated.

This chapter details essential laboratory procedures for proteomic biomarker discovery. It covers sample preparation, including protein extraction, digestion, and fractionation, for accurate mass spectrometry analysis.

Keywords:
Biomarker discoveryIn-gel digestion of proteinsIn-solution digestion of proteinsLC-MS/MSLTQ OrbitrapMass spectrometryProteomicsSEQUESTSilver stain

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Area of Science:

  • Proteomics
  • Biomarker Discovery
  • Analytical Chemistry

Background:

  • Identifying protein biomarkers in the proteome is crucial but challenging.
  • Mass spectrometry (MS) offers sensitive and accurate protein identification.
  • Sample preparation, including extraction and fractionation, is vital for MS-based biomarker discovery.

Purpose of the Study:

  • To provide practical laboratory protocols for proteomic sample preparation.
  • To guide researchers through digestion, reduction, alkylation, and cleanup steps.
  • To facilitate biomarker discovery using mass spectrometry.

Main Methods:

  • Detailed procedures for protein extraction from biological samples.
  • Step-by-step protocols for enzymatic protein digestion.
  • Methods for protein fractionation to simplify complex mixtures.
  • Sample cleanup techniques prior to mass spectrometry analysis.

Main Results:

  • The chapter offers a comprehensive guide to essential sample preparation techniques.
  • Standardized protocols ensure reproducibility in proteomic workflows.
  • The methods described are applicable to various biological matrices.

Conclusions:

  • Efficient sample preparation is key to successful proteomic biomarker discovery.
  • Practical laboratory procedures enhance the accuracy and sensitivity of mass spectrometry.
  • This chapter serves as a valuable resource for researchers in the field.