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

Epitope tagging and protein surveillance.

P A Kolodziej, R A Young

    Methods in Enzymology
    |January 1, 1991
    PubMed
    Summary

    Epitope tagging offers a precise and economical method for protein research, overcoming limitations of traditional antibodies. This technique ensures high-affinity, specific antibody binding for accurate protein purification and complex analysis.

    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

    Management strategies following recurrence of resected retroperitoneal sarcoma: A 10-year observational study.

    European journal of surgical oncology : the journal of the European Society of Surgical Oncology and the British Association of Surgical Oncology·2025
    Same author

    APOBEC signature mutation generates an oncogenic enhancer that drives LMO1 expression in T-ALL.

    Leukemia·2017
    Same author

    Identification of a cancer stem cell-specific function for the histone deacetylases, HDAC1 and HDAC7, in breast and ovarian cancer.

    Oncogene·2016
    Same author

    TRIB2 reinforces the oncogenic transcriptional program controlled by the TAL1 complex in T-cell acute lymphoblastic leukemia.

    Leukemia·2015
    Same author

    The lived experience of visual creative expression for young adult cancer survivors.

    European journal of cancer care·2014
    Same author

    Career development of adolescents: An ecological perspective.

    Journal of youth and adolescence·2013

    Area of Science:

    • Molecular Biology
    • Biochemistry
    • Yeast Genetics

    Background:

    • Traditional antibody-based protein studies face challenges with antibody affinity, specificity, and abundance.
    • Direct antibody generation may not always yield optimal reagents for protein research.

    Purpose of the Study:

    • To highlight the advantages of epitope tagging over direct antibody use for protein research.
    • To demonstrate the efficacy of epitope tagging in yeast for validating protein function and enabling purification.

    Main Methods:

    • Oligonucleotide-mediated site-directed mutagenesis to add epitope coding sequences to genes.
    • Complementation testing in yeast to assess in vivo function of modified proteins.
    • Immunoprecipitation and affinity purification using monoclonal antibodies against unique epitopes.

    Main Results:

    • Epitope tagging provides universal, economical, and precise protein modification.
    • Yeast complementation assays readily confirm the in vivo functionality of tagged proteins.
    • High-affinity, specific, and abundant antibody binding is achieved through unique epitope recognition.
    • Purification of proteins, even those defective in essential functions, is facilitated.
    • Protein complexes can be accurately defined by identifying immunoprecipitable polypeptides specific to the tagged protein.

    Conclusions:

    • Epitope tagging is a superior strategy for protein research, offering significant advantages in reagent quality and experimental flexibility.
    • The method allows for robust protein purification, functional studies, and complex analysis, even for challenging targets.
    • This approach enhances the precision and efficiency of molecular and biochemical investigations.

    Related Experiment Videos