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Related Experiment Video

Updated: May 30, 2026

Proteomic Profiling of Macrophages by 2D Electrophoresis
07:53

Proteomic Profiling of Macrophages by 2D Electrophoresis

Published on: November 4, 2014

Proteomic biosignatures for monocyte-macrophage differentiation.

Stephanie D Kraft-Terry1, Howard E Gendelman

  • 1Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE 68198-5880, USA.

Cellular Immunology
|July 27, 2011
PubMed
Summary

Pulsed stable isotope labeling of amino acids in cell culture (pSILAC) tracked protein changes during monocyte-macrophage differentiation. This revealed key proteins involved in cell homeostasis and morphology during transformation.

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Area of Science:

  • Proteomics
  • Cell Biology
  • Biochemistry

Background:

  • Monocyte-macrophage differentiation is a critical process in immunity and inflammation.
  • Understanding protein dynamics during this transformation is essential for elucidating cellular functions.
  • Previous studies have lacked detailed temporal analysis of protein synthesis during this process.

Purpose of the Study:

  • To investigate protein dynamics and de novo protein production during monocyte-macrophage differentiation using pulsed stable isotope labeling.
  • To identify specific proteins and pathways involved in monocyte transformation.
  • To provide a temporal map of protein synthesis from 3 to 7 days of differentiation.

Main Methods:

  • Pulsed stable isotope labeling of amino acids in cell culture (pSILAC) was employed for metabolic labeling of newly synthesized proteins.

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Isolation of Human Monocytes by Double Gradient Centrifugation and Their Differentiation to Macrophages in Teflon-coated Cell Culture Bags
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Isolation of Human Monocytes by Double Gradient Centrifugation and Their Differentiation to Macrophages in Teflon-coated Cell Culture Bags

Published on: September 9, 2014

Proteomic Analysis of Human Macrophage Polarization Under a Low Oxygen Environment
10:15

Proteomic Analysis of Human Macrophage Polarization Under a Low Oxygen Environment

Published on: January 7, 2019

Related Experiment Videos

Last Updated: May 30, 2026

Proteomic Profiling of Macrophages by 2D Electrophoresis
07:53

Proteomic Profiling of Macrophages by 2D Electrophoresis

Published on: November 4, 2014

Isolation of Human Monocytes by Double Gradient Centrifugation and Their Differentiation to Macrophages in Teflon-coated Cell Culture Bags
09:32

Isolation of Human Monocytes by Double Gradient Centrifugation and Their Differentiation to Macrophages in Teflon-coated Cell Culture Bags

Published on: September 9, 2014

Proteomic Analysis of Human Macrophage Polarization Under a Low Oxygen Environment
10:15

Proteomic Analysis of Human Macrophage Polarization Under a Low Oxygen Environment

Published on: January 7, 2019

  • Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used for protein identification.
  • MaxQuant software was utilized for quantitative analysis of protein levels.
  • Ingenuity Pathway Analysis (IPA) was performed to assess protein-protein interactions and biological pathways.

    • Main Results:

    • pSILAC successfully quantified de novo protein production over a 3 to 7-day culture period.
    • Identified proteins were significantly associated with cell homeostasis, free radical scavenging, molecular protein transport, carbohydrate metabolism, small molecule chemistry, and cell morphology.
    • The study revealed specific protein dynamics linked to the transformation of monocytes into macrophages.

    Conclusions:

    • pSILAC is a powerful technique for studying protein dynamics during cellular differentiation.
    • The identified protein networks provide insights into the molecular mechanisms governing monocyte-macrophage differentiation.
    • These findings contribute to a better understanding of cellular transformation and its associated biological events.