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Related Concept Videos

Size-Exclusion Chromatography01:08

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In size-exclusion chromatography (SEC), also known as molecular-exclusion or gel-permeation chromatography, molecules are separated based on their sizes. This technique is important for separating large molecules such as polymers and biomolecules. The two classes of micron-sized stationary phases encountered in SEC are silica particles and cross-linked polymer resin beads. Both materials are porous, but their pore sizes vary significantly.
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In inductively coupled plasma–mass spectrometry (ICP–MS), an inductively coupled plasma (ICP) torch is used as an atomizer and ionizer. Solid samples are dissolved and volatilized before being introduced into the high-temperature argon plasma, while solution samples are nebulized and passed through the high-temperature argon plasma. Plasma dissociates the analytes and ionizes their component atoms to form a mixture of positive ions and molecular species. The positive ions are then...
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Related Experiment Video

Updated: Mar 21, 2026

Standards for Quantitative Metalloproteomic Analysis Using Size Exclusion ICP-MS
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Standards for Quantitative Metalloproteomic Analysis Using Size Exclusion ICP-MS

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Standards for Quantitative Metalloproteomic Analysis Using Size Exclusion ICP-MS.

Amber Lothian1, Blaine R Roberts2

  • 1Metalloproteomics Laboratory, Neuroproteomics Facility, The Florey Institute of Neuroscience and Mental Health.

Journal of Visualized Experiments : Jove
|May 12, 2016
PubMed
Summary

This study introduces a new method to quantify metalloproteins in complex biological samples. The technique uses size exclusion chromatography coupled with inductively coupled plasma - mass spectrometry (SEC-ICP-MS) to measure metalloprotein levels, aiding disease research.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Neuroscience

Background:

  • Metals are crucial cofactors for protein function.
  • Disrupted metal homeostasis is linked to neurodegenerative diseases like Alzheimer's and Parkinson's.
  • Quantifying metalloproteins in complex biological samples presents significant challenges.

Purpose of the Study:

  • To develop and validate a novel method for quantifying metalloproteins in complex samples.
  • To establish a technique for comparing metalloprotein abundances across different biological specimens.
  • To facilitate research into the role of metalloproteins in neurodegenerative diseases.

Main Methods:

  • Utilized size exclusion chromatography coupled with inductively coupled plasma - mass spectrometry (SEC-ICP-MS).
  • Calibrated the size exclusion column using six known metalloprotein standards.
  • Quantified trace elements (iron, copper, zinc, cobalt, iodine) for metalloprotein measurement.

Main Results:

  • Successfully quantified metalloproteins in complex human brain and plasma samples.
  • Demonstrated the ability to quantitatively compare metalloprotein abundances.
  • Established SEC-ICP-MS as a viable technique for metalloprotein analysis.

Conclusions:

  • The SEC-ICP-MS method provides a robust approach for metalloprotein quantification.
  • This technique enables quantitative comparisons of metalloprotein levels between biological samples.
  • The method holds promise for elucidating the role of metalloproteins in neurodegenerative and other diseases.