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

Post-Electrophoretic Identification of Oxidized Proteins.

Craig C Conrad1, John M Talent, Christina A Malakowsky

  • 1Molecular Aging Unit, Department of Molecular Biology and Immunology. Molecular Aging Unit, Department of Molecular Biology and Immunology, University of North Texas Health Science Center, Fort Worth, Texas, 76107. USA.Office of Research and Biotechnology. ME-2 Room 806, 3500 Camp Bowie Blvd, Fort Worth, Texas 76107. USA. rgracy@hsc.unt.edu

Biological Procedures Online
|May 8, 2003
PubMed
Summary

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

Protection of HT22 neuronal cells against glutamate toxicity mediated by the antioxidant activity of Pueraria candollei var. mirifica extracts.

Journal of natural medicines·2010
Same author

Decreasing photobleaching by silver nanoparticles on metal surfaces: application to muscle myofibrils.

Journal of biomedical optics·2008
Same author

Proteomic identification of specific oxidized proteins in ApoE-knockout mice: relevance to Alzheimer's disease.

Free radical biology & medicine·2004
Same author

Anti-apoptotic proteins are oxidized by Abeta25-35 in Alzheimer's fibroblasts.

Biochimica et biophysica acta·2003
Same author

Vitamin E prevents oxidation of antiapoptotic proteins in neuronal cells.

Proteomics·2003
Same author

Flavones from Scutellaria baicalensis Georgi attenuate apoptosis and protein oxidation in neuronal cell lines.

Biochimica et biophysica acta·2002

Identifying proteins susceptible to oxidative modification is challenging. This new method uses electrophoresis and blotting to quantify protein oxidation, aiding disease research.

Area of Science:

  • Biochemistry
  • Proteomics
  • Analytical Chemistry

Background:

  • Oxidative modification of proteins is implicated in numerous human diseases.
  • Identifying specific, susceptible proteins is analytically challenging.

Purpose of the Study:

  • To develop a method for identifying proteins susceptible to oxidative modification.
  • To quantify the specific oxidation index of individual proteins.

Main Methods:

  • Proteins separated by polyacrylamide gel electrophoresis (PAGE).
  • Electroblotting of underivatized proteins onto PVDF membranes.
  • Derivatization of carbonyls with 2,4-DNP, followed by immunostaining and colloidal gold protein staining.

Main Results:

Related Experiment Videos

  • A method enabling quantification of both protein and oxidized protein levels.
  • Determination of the specific oxidation index for individual proteins.
  • Established optimal staining sequence: electrophoresis, electroblotting, derivatization, immunostaining, and protein staining.

    • Conclusions:

    • The developed method allows for the identification and quantification of oxidatively modified proteins.
    • This technique facilitates the study of protein oxidation's role in human diseases.
    • The method provides a tool for determining the specific oxidation index of proteins.