Elispot Analysis
Elispot Analysis - Related Videos
11:53
ELISPOT Assay: Detection of IFN-γ Secreting Splenocytes
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2023
Source: Tonya J. Webb1 1 Department of Microbiology and Immunology, University of Maryland School of Medicine and the Marlene and Stewart Greenebaum Comprehensive Cancer Center, Baltimore, Maryland 21201 ELISPOT is a standardized, reproducible assay used to detect cellular immune responses. The assay utilizes an enzyme-linked immunosorbent assay (ELISA)- based method to detect single-cell immune responses which can be visualized by spots, hence the name ELISPOT. ELISPOT was first described in...
04:20
Functional Assessment of Antibody-Secreting B Cells Using ELISpot Assay
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2025
This video demonstrates the use of the ELISpot assay to assess the function of antibody-secreting B cells, ASCs. The antibodies secreted by the ASCs interact with the membrane-bound polyclonal antibody fragments, which are detected by the enzyme-conjugated IgG or IgM antibodies. The colored spot produced in each well indicates the presence of the specific antibody's isotype, which correlates with the B cells' secretory activity.
08:26
The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity
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Cited by 10 •
2016
Human peripheral blood is commonly used for the assessment of the humoral immune response. Here, the methods for isolating human B cells from peripheral blood, differentiating human B cells into antibody (Ab)-secreting B cells (ASCs) in culture, and enumerating the total IgM- and IgG-ASCs via an ELISpot assay are described.
06:13
Enzyme-linked Immunospot Assay (ELISPOT): Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens
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Cited by 3 •
2010
Identification of microbial targets of adaptive immunity in idiopathic diseases can be accomplished by the use of the enzyme-linked immunospot assay.
08:13
Optimized Interferon-gamma ELISpot Assay to Measure T Cell Responses in the Guinea Pig Model after Vaccination
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Cited by 3 •
2019
The development of the interferon-gamma ELISpot assay for guinea pig PBMCs allows the characterization of T-cell responses in this highly relevant model for studying infectious diseases. We have applied the assay to measure T cell responses associated with intradermal delivery of DNA vaccines.
FAQs
The capture antibody anchors the assay’s spatial resolution: molecules released by a cell are retained close to that cell on the coated membrane rather than dispersing through the culture. A labeled detection antibody then reveals these localized capture events. This arrangement makes secretory activity visible as discrete signals suitable for counting.
Spot enumeration provides a frequency-based readout rather than only a bulk measurement of total secreted material. Because signals remain associated with localized secretion events, the assay can identify how many cells responded within the tested population. This single-cell perspective is especially useful when a response is present in only a small fraction of cells.
The measured target determines which immune response is profiled. By detecting a secreted cytokine or antibody, Elispot Analysis can support evaluation of antigen-specific T-cell or B-cell responses in studies of infection and vaccination. This target-specific design also makes the assay useful for immune monitoring, where response type matters alongside responding-cell frequency.
A basic workflow starts by placing cells onto a membrane coated with a capture antibody and culturing them so secreted molecules bind nearby. After secretion, labeled antibodies detect the captured material, producing visible spots. The resulting spot pattern can then be interpreted as evidence of responding cells and compared across experimental immune conditions.
Elispot Analysis is useful when investigators need to detect antigen-specific responses with high sensitivity and single-cell resolution. In infection studies, it can evaluate host responses to pathogen exposure; in vaccination studies, it can assess vaccine-associated immune responses. The same readout also supports immune monitoring when response magnitude or responding-cell frequency is important.
Results can indicate whether a tested population contains cells that respond to a defined antigen by secreting the measured cytokine or antibody. This supports comparisons of host immunity across infection or vaccination settings and can extend to therapeutic-response monitoring. Interpretation should focus on responding-cell frequency represented by spots, while the secreted target identifies response type.