Cell-based ELISpot protocol to detect and quantify antigen-specific antibody-secreting cells in murine whole-organ single-cell suspensions

Antibody-secreting cells (ASCs) are the hallmark of the adaptive immune response to vaccines or infections. Here, we present a protocol to quantify the number of ASCs using an enzyme-linked immunospot assay (ELISpot). We describe the preparation of cell samples, ELISpot plates, and their quantification. This assay has potential applications in vaccine studies and research on the adaptive immune system or ASC-associated diseases. For complete details on the use and execution of this protocol please, refer to Bierling et al.<sup>1</sup>.

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Publication Details

Journal
STAR Protocols
Published
2025-07-16
DOI
https://doi.org/10.1016/j.xpro.2025.103797
Primary Topic
Monoclonal and Polyclonal Antibodies Research
Type
article
Field-Weighted Citation Impact
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article

Cell-based ELISpot protocol to detect and quantify antigen-specific antibody-secreting cells in murine whole-organ single-cell suspensions

Katharina Pracht, Stephan Menzel, Christina Brunner, Franziska Rost et al.
STAR Protocols
Monoclonal and Polyclonal Antibodies Research
article

Cell-based ELISpot protocol to detect and quantify antigen-specific antibody-secreting cells in murine whole-organ single-cell suspensions

Katharina Pracht, Stephan Menzel, Christina Brunner, Franziska Rost, Joana Côrte‐Real
article en

Abstract

Antibody-secreting cells (ASCs) are the hallmark of the adaptive immune response to vaccines or infections. Here, we present a protocol to quantify the number of ASCs using an enzyme-linked immunospot assay (ELISpot). We describe the preparation of cell samples, ELISpot plates, and their quantification. This assay has potential applications in vaccine studies and research on the adaptive immune system or ASC-associated diseases. For complete details on the use and execution of this protocol please, refer to Bierling et al.<sup>1</sup>.

STAR ProtocolsVol. 6(3)
Friedrich-Alexander-Universität Erlangen-Nürnberg (DE), Universitätsklinikum Erlangen (DE)
Openalex Percentile: Top 74%
Monoclonal and Polyclonal Antibodies Research
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