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Related Concept Videos

Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

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...
Peptide Identification Using Tandem Mass Spectrometry01:33

Peptide Identification Using Tandem Mass Spectrometry

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.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
MALDI-TOF Mass Spectrometry01:19

MALDI-TOF Mass Spectrometry

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...
Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

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.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
Mass Spectrometry: Overview01:19

Mass Spectrometry: Overview

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...
Mass Spectrometers01:16

Mass Spectrometers

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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Low Molecular Weight Protein Enrichment on Mesoporous Silica Thin Films for Biomarker Discovery
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Low Molecular Weight Protein Enrichment on Mesoporous Silica Thin Films for Biomarker Discovery

Published on: April 17, 2012

Mass spectrometry-based biomarker discovery.

Weidong Zhou1, Emanuel F Petricoin, Caterina Longo

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

Methods in Molecular Biology (Clifton, N.J.)
|November 15, 2011
PubMed
Summary
This summary is machine-generated.

This chapter details essential laboratory procedures for sample preparation in proteomic biomarker discovery. It focuses on protein digestion and fractionation techniques crucial for mass spectrometry analysis.

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Navigating the Mass Spectrometry-Based Proteomic Data Using Free Computational Tools
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Area of Science:

  • Proteomics
  • Biomarker Discovery
  • Analytical Chemistry

Background:

  • Identifying protein biomarkers in the proteome is a critical yet challenging goal in biomedical research.
  • Mass spectrometry (MS) is a powerful technology for protein identification and quantification, essential for biomarker discovery.
  • Effective sample preparation, including protein extraction, digestion, and fractionation, is a prerequisite for successful MS-based proteomic analysis.

Purpose of the Study:

  • To provide practical laboratory protocols for sample digestion and protein fractionation.
  • To outline key steps in the workflow for mass spectrometry-based biomarker discovery.
  • To facilitate the accurate and sensitive assessment of proteins for identifying candidate biomarkers.

Main Methods:

  • Detailed procedures for enzymatic or chemical digestion of proteins from various biological samples.
  • Methods for protein fractionation using techniques such as liquid chromatography (e.g., HPLC, FPLC) or gel electrophoresis.
  • Best practices for sample handling and preparation to ensure integrity and minimize loss.

Main Results:

  • Standardized protocols for reproducible protein digestion and fractionation.
  • Optimized workflows enabling efficient sample processing for subsequent mass spectrometry analysis.
  • Guidelines for achieving high-quality data crucial for biomarker identification.

Conclusions:

  • Robust sample digestion and fractionation are fundamental for successful proteomic biomarker discovery using mass spectrometry.
  • The provided practical procedures aim to enhance the efficiency and reliability of proteomic workflows.
  • Adherence to these methods will aid researchers in advancing the field of biomarker identification.