Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Enzyme-Linked Immunosorbent Assay

Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or...

You might also read

Related Articles

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

Sort by

Same author

Myeloid-derived suppressor cell (MDSC)-like neutrophils induced by pulmonary infection with <i>Coccidioides posadasii</i> exacerbate disease by suppressing CD4<sup>+</sup> T cell immunity.

mBio·2026

Same author

Editorial for exploring understudied inflammatory diseases.

Expert review of clinical immunology·2026

Same author

MALDI-TOF mass spectrometry imaging of sulfatide lipid expression in the CNS of mice with experimental autoimmune encephalomyelitis.

Scientific reports·2026

Same author

C/EBPβ-induced alternative splicing of RCAN1 generates a potent TCR-T target in mesenchymal glioblastoma.

Cellular & molecular immunology·2025

Same author

A Double-Humanized Murine Model in Bladder Cancer: A Novel Preclinical Model for Cancer Immunology Research.

Cancer medicine·2025

Same author

CD56 on intratumoral NK cells: orchestrating NK cell-mediated anti-tumor effects in bladder cancer.

Neoplasia (New York, N.Y.)·2025

Search research articles

Related Experiment Video

Updated: Apr 25, 2026

Enzyme-linked Immunospot Assay ELISPOT: Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens

Enzyme-linked Immunospot Assay ELISPOT: Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens

Published on: November 23, 2010

ELISPOT Techniques.

Niannian Ji¹, Thomas G Forsthuber

¹Department of Biology, The University of Texas at San Antonio, San Antonio, TX, USA.

Methods in Molecular Biology (Clifton, N.J.)

|August 14, 2014

Summary

The enzyme-linked immunospot (ELISPOT) assay is a key method for detecting cytokine-producing immune cells. This guide provides optimized protocols and troubleshooting for ELISPOT assays in human and mouse samples.

Area of Science:

Immunology
Cellular Immunology
Assay Development

Background:

The enzyme-linked immunospot (ELISPOT) assay is a crucial technique for enumerating antigen-specific cytokine-producing or antibody-secreting immune cells.
It serves as a highly sensitive immunological and diagnostic tool for detecting and quantifying low-frequency immune cells in various human and animal tissues.

Purpose of the Study:

To provide practical, optimized protocols for cytokine ELISPOT assays.
To detail methods for detecting mouse and human cytokine-producing immune cells, including peripheral blood mononuclear cells (PBMC).
To offer troubleshooting and optimization strategies for challenging tissue samples.

Main Methods:

Utilizes pairs of capture and detection antibodies to identify cytokine release sites.

More Related Videos

The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity

The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity

Published on: December 14, 2016

Development of an IFN-γ ELISpot Assay to Assess Varicella-Zoster Virus-specific Cell-mediated Immunity Following Umbilical Cord Blood Transplantation

Development of an IFN-γ ELISpot Assay to Assess Varicella-Zoster Virus-specific Cell-mediated Immunity Following Umbilical Cord Blood Transplantation

Published on: July 9, 2014

Related Experiment Videos

Last Updated: Apr 25, 2026

Enzyme-linked Immunospot Assay ELISPOT: Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens

Enzyme-linked Immunospot Assay ELISPOT: Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens

Published on: November 23, 2010

The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity

The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity

Published on: December 14, 2016

Development of an IFN-γ ELISpot Assay to Assess Varicella-Zoster Virus-specific Cell-mediated Immunity Following Umbilical Cord Blood Transplantation

Development of an IFN-γ ELISpot Assay to Assess Varicella-Zoster Virus-specific Cell-mediated Immunity Following Umbilical Cord Blood Transplantation

Published on: July 9, 2014

Focuses on optimized conditions for spot formation by immune cells (e.g., T cells).

Employs established ELISPOT assay principles for immune cell enumeration.

Main Results:

The study details optimized protocols for cytokine ELISPOT assays.
Provides practical guidance for detecting cytokine-producing immune cells in mouse and human samples.
Includes troubleshooting tips for problematic samples, enhancing assay reliability.

Conclusions:

Optimized ELISPOT protocols enhance the detection and quantification of cytokine-producing immune cells.
The assay is effective for analyzing immune responses in various biological samples.
This work facilitates more robust immune cell analysis in research and diagnostics.