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 Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Generating Monoclonal Antibodies.

Cold Spring Harbor protocols·2022
Same author

Determining the Class and Subclass of Monoclonal Antibodies Using Antibody Capture on Anti-Immunoglobulin Antibody-Coated Plates.

Cold Spring Harbor protocols·2022
Same author

Immunizing Animals.

Cold Spring Harbor protocols·2022
Same author

Hybridoma Screening by Antibody Capture: Flow Cytometry/FACS with Permeabilized Cells to Detect Intracellular Binding.

Cold Spring Harbor protocols·2022
Same author

Hybridoma Screening by Antibody Capture: Flow Cytometry/FACS with Whole Cells to Detect Cell-Surface Binding.

Cold Spring Harbor protocols·2022
Same author

Hybridoma Screening by Antibody Capture: Enzyme-Linked Detection (Indirect ELISA) in Polyvinyl Chloride Wells.

Cold Spring Harbor protocols·2022

Related Experiment Video

Updated: Jan 2, 2026

Murine Model of Epicutaneously-Induced Immunomodulation
09:07

Murine Model of Epicutaneously-Induced Immunomodulation

Published on: June 24, 2025

395

Immunizing Mice and Rats with Nitrocellulose-Bound Antigen.

Edward A Greenfield

    Cold Spring Harbor Protocols
    |December 4, 2019
    PubMed
    Summary

    This study presents a novel method for antigen purification and immunization using nitrocellulose membranes. Immobilizing protein antigens on nitrocellulose enhances immunogenicity and simplifies delivery, improving antigen-specific immune responses.

    Area of Science:

    • Biochemistry
    • Immunology
    • Protein Chemistry

    Background:

    • Gel electrophoresis is a common method for separating target antigens from complex protein mixtures like cell lysates.
    • Extracting antigens from gel slices and eluting them is inefficient, leading to protein loss and potential animal intolerance to injected gel pieces.

    Purpose of the Study:

    • To develop an improved method for antigen purification and immunization.
    • To enhance the immunogenicity of protein antigens for antibody production.

    Main Methods:

    • Protein antigens were separated by gel electrophoresis and transferred onto nitrocellulose membranes.
    • Antigens immobilized on nitrocellulose were injected subcutaneously or intraperitoneally, or implanted subdermally.
    • The slow release of antigens from nitrocellulose was hypothesized to act as an adjuvant.

    More Related Videos

    A Rapid, Simple, and Standardized Homogenization Method to Prepare Antigen/Adjuvant Emulsions for Inducing Experimental Autoimmune Encephalomyelitis
    05:44

    A Rapid, Simple, and Standardized Homogenization Method to Prepare Antigen/Adjuvant Emulsions for Inducing Experimental Autoimmune Encephalomyelitis

    Published on: December 9, 2022

    4.3K
    Rat Model of Widespread Cerebral Cortical Demyelination Induced by an Intracerebral Injection of Pro-Inflammatory Cytokines
    09:46

    Rat Model of Widespread Cerebral Cortical Demyelination Induced by an Intracerebral Injection of Pro-Inflammatory Cytokines

    Published on: September 21, 2021

    5.1K

    Related Experiment Videos

    Last Updated: Jan 2, 2026

    Murine Model of Epicutaneously-Induced Immunomodulation
    09:07

    Murine Model of Epicutaneously-Induced Immunomodulation

    Published on: June 24, 2025

    395
    A Rapid, Simple, and Standardized Homogenization Method to Prepare Antigen/Adjuvant Emulsions for Inducing Experimental Autoimmune Encephalomyelitis
    05:44

    A Rapid, Simple, and Standardized Homogenization Method to Prepare Antigen/Adjuvant Emulsions for Inducing Experimental Autoimmune Encephalomyelitis

    Published on: December 9, 2022

    4.3K
    Rat Model of Widespread Cerebral Cortical Demyelination Induced by an Intracerebral Injection of Pro-Inflammatory Cytokines
    09:46

    Rat Model of Widespread Cerebral Cortical Demyelination Induced by an Intracerebral Injection of Pro-Inflammatory Cytokines

    Published on: September 21, 2021

    5.1K

    Main Results:

    • Nitrocellulose-immobilized protein antigens demonstrated enhanced immunogenicity in some cases.
    • This method simplifies antigen delivery and potentially improves immune responses compared to direct gel injection.

    Conclusions:

    • Immobilizing protein antigens on nitrocellulose offers a viable alternative for antigen purification and immunization.
    • This technique can improve antigen immunogenicity and simplify the immunization process for research applications.