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 Videos

Quantitative and reproducible two-dimensional gel analysis using Phoretix 2D Full.

P Mahon1, P Dupree

  • 1University of Cambridge, Department of Biochemistry, United Kingdom.

Electrophoresis
|July 24, 2001
PubMed
Summary

Phoretix 2D Full software provides reliable protein quantification using colloidal Coomassie staining in two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). User seeding improves data reproducibility, while background subtraction algorithms reduce it.

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

Impact of precision oncology research in pediatric poor prognosis cancer: patient, parent and healthcare provider perspectives.

BMC cancer·2026
Same author

Interaction of a glucuronoyl esterase with complex fragments of the plant cell wall hemicellulose.

International journal of biological macromolecules·2026
Same author

Continuous spinal anaesthesia in frail patients for hip fracture surgery.

Irish medical journal·2025
Same author

The structure of EXTL3 helps to explain the different roles of bi-domain exostosins in heparan sulfate synthesis.

Nature communications·2022
Same author

Placental volume at 11 weeks is associated with offspring bone mass at birth and in later childhood: Findings from the Southampton Women's Survey.

Placenta·2020
Same author

Fascial layers influence the spread of injectate during ultrasound-guided infraclavicular brachial plexus block: a cadaver study.

British journal of anaesthesia·2018

Area of Science:

  • Proteomics
  • Biochemistry
  • Molecular Biology

Background:

  • Quantitative analysis of protein expression relies on accurate 2-D PAGE.
  • Reliable 2-D gel image analysis requires software with a wide dynamic range and linear response.
  • Colloidal Coomassie Brilliant Blue G-250 is a common stain for visualizing proteins on 2-D gels.

Purpose of the Study:

  • To evaluate the linearity and quantitative accuracy of Phoretix 2D Full software for 2-D gel image analysis.
  • To assess the impact of specific analysis parameters on data reproducibility and accuracy.
  • To validate the software's performance using a complex plant protein mixture.

Main Methods:

  • Quantitative two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was performed.

Related Experiment Videos

  • Gels were stained with colloidal Coomassie Brilliant Blue G-250.
  • Images were analyzed using Phoretix 2D Full software, with and without background subtraction and user seeding.

    • Main Results:

    • Phoretix 2D Full demonstrated a linear response for protein quantification over three orders of magnitude.
    • The software accurately measured protein quantity changes in a complex mixture of Arabidopsis thaliana proteins.
    • User seeding significantly improved data reproducibility, whereas background subtraction algorithms decreased it.

    Conclusions:

    • Phoretix 2D Full software provides a linear and quantitative measure of protein levels from 2-D gels stained with colloidal Coomassie.
    • Careful selection of analysis parameters, such as user seeding, is crucial for reproducible proteomic data.
    • The findings support the use of Phoretix 2D Full for reliable protein expression analysis in complex biological samples.