Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Video

Updated: May 23, 2026

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
10:37

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Published on: November 15, 2017

Isobaric labeling and data normalization without requiring protein quantitation.

Phillip D Kim1, Bhavinkumar B Patel, Anthony T Yeung

  • 1Developmental Therapeutics Program, Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111-2497, USA.

Journal of Biomolecular Techniques : JBT
|April 3, 2012
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Challenges in Spinal Endoscopy.

World neurosurgery·2022
Same author

Retrospective analysis of accuracy and positive predictive value of preoperative lumbar MRI grading after successful outcome following outpatient endoscopic decompression for lumbar foraminal and lateral recess stenosis.

Clinical neurology and neurosurgery·2019
Same author

The State of the Art in Minimally Invasive Spine Surgery.

BioMed research international·2017
Same author

Haploinsufficiency in tumor predisposition syndromes: altered genomic transcription in morphologically normal cells heterozygous for VHL or TSC mutation.

Oncotarget·2016
Same author

The learning curve in foraminal endoscopic discectomy: experience needed to achieve a 90% success rate.

SAS journal·2015
Same author

The Evolution and Advancement of Endoscopic Foraminal Surgery: One Surgeon's Experience Incorporating Adjunctive Techologies.

SAS journal·2015

A new method called exponentially modified protein abundance index (emPAI)-MW deconvolution (EMMOL) normalizes isobaric tag for relative and absolute quantification (iTRAQ) data. This allows comparing many samples, even with minute protein amounts, without a reference standard.

Area of Science:

  • Proteomics
  • Quantitative Mass Spectrometry

Background:

  • Isobaric multiplexed quantitative proteomics is valuable for high-resolution sample isolation.
  • Accurate normalization of isobaric reporter ratios is crucial for reliable quantitative proteomics, especially when dealing with limited sample quantities.

Purpose of the Study:

  • To introduce a novel workflow, exponentially modified protein abundance index (emPAI)-MW deconvolution (EMMOL), for normalizing isobaric reporter ratios.
  • To enable quantitative comparison of multiple samples without a reference standard, particularly when using small or unknown protein amounts.
  • To optimize sample preparation for minute protein quantities in quantitative proteomics experiments.

Main Methods:

  • Developed and applied the EMMOL workflow for deconvolution of isobaric tags for relative and absolute quantification (iTRAQ) data.
Keywords:
EMMOLLCMiTRAQoptimizationproteome

More Related Videos

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level
08:29

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level

Published on: April 19, 2019

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling
11:53

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling

Published on: July 1, 2014

Related Experiment Videos

Last Updated: May 23, 2026

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
10:37

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Published on: November 15, 2017

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level
08:29

Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level

Published on: April 19, 2019

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling
11:53

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling

Published on: July 1, 2014

  • Utilized emPAI, target protein molecular weight (MW), and iTRAQ reporter ratios to calculate protein quantities.
  • Implemented an optimized iTRAQ labeling protocol starting with as little as 5 microg of protein.

    • Main Results:

    • EMMOL successfully normalized iTRAQ data, yielding protein quantities for each sample without prior knowledge or a reference standard.
    • Demonstrated EMMOL's capability by comparing four pooled samples with 20-fold differences in protein abundance, showing no compression of expression ratios.
    • Validated the analysis of minute samples with an optimized iTRAQ labeling protocol.

    Conclusions:

    • EMMOL provides a robust method for normalizing isobaric quantitative proteomics data, enabling accurate comparisons across experiments and samples.
    • The workflow facilitates the analysis of limited biological samples, expanding the applicability of quantitative proteomics.
    • EMMOL allows for the comparison of proteomes from separate iTRAQ experiments without the need for a common reference sample.