ELISpot and FluoroSpot

Quantification of immune responses at the single-cell level

Mabtech is a privately owned Swedish biotech company that emerged from the Department of Immunology at Stockholm University in 1986.

Mabtech develops and manufactures high quality monoclonal antibodies and kits for ELISA, ELISpot and FluoroSpot and is a world leader in advancing the ELISpot and FluoroSpot methods.

Learn more about ELISpot and FluoroSpot Applications.

ELISpot

ELISpot is a robust and straightforward assay for detecting and quantifying antigen-specific immune responses at the single-cell level.

The ELISpot assay captures the presence of cytokines immediately after secretion, in contrast to measurements that are skewed by receptor binding or protease degradation. With detection levels as low as one cell in 250,000, ELISpot is one of the most sensitive cellular assays. It is carried out in a 96-well plate, and an automated ELISpot reader is used for analysis. The assay is therefore easy to perform and allows rapid analysis of a large number of samples.

ELISpot in Summary:

•  Sensitive immunoassay
•  Measures secretion of cytokines
•  Easy to perform
•  Robust and easy to standardize
•  Suitable for large-scale trials also in field settings

FIG: Schematic illustration of the principle of the ELISpot assay

ELISpot from Mabtech means quality through and through. You’re getting more than a kit, you’re getting years of assay innovation, optimized protocols, and the tools you need to generate data you can trust to drive your research and therapies forward.

• Decades of expertise: 40 years dedicated to advancing ELISpot technology, with trusted antibodies and kits used in thousands of peer-reviewed studies and clinical trials.
• Exceptional sensitivity: Detects even the rarest antigen-specific T and B cells, ensuring no important response goes undetected.
• In-house antibody development: Proprietary monoclonal antibodies give unmatched specificity and affinity, batch after batch.
• Integrated reading solutions: Mabtech ASTOR™ 2 and IRIS™ 2 readers with the patented RAWspot™ algorithm provide the highest precision in spot detection and counting available.

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FluoroSpot

FluoroSpot is the multiplex version of ELISpot. Just like in ELISpot, target proteins secreted by cells are captured by specific antibodies immediately after secretion and throughout the stimulation, making it an extremely sensitive sandwich assay. The difference to ELISpot lies in the mode of detection: where ELISpot relies on an enzymatic reaction, FluoroSpot utilizes fluorescence.

FluoroSpot from Mabtech means quality through and through. You’re getting more than a kit, you’re getting years of assay innovation, optimized protocols, and the tools you need to generate data you can trust to drive your research and therapies forward.

• Decades of expertise: Mabtech's first FluoroSpot kits were launched in 2008, and ever since, they’ve been advancing the technology, with trusted antibodies and kits used in hundreds of peer-reviewed studies and clinical trials.
• Exceptional sensitivity: FluoroSpot is hundreds of times more sensitive than intracellular staining in flow cytometry. Detects even the rarest antigen-specific T and B cells, ensuring no important response goes undetected.
• Polyfunctional assessment: The multiplexing capacity of FluoroSpot allows for evaluating cells secreting more than one analyte, giving even more detailed immune insights.
• Relative spot volume: The patented RAWspot™ algorithm allows for measurement of the relative amount of analyte of interest secreted, adding more nuanced insights in your immune profiling.
• In-house antibody development: Proprietary monoclonal antibodies give unmatched specificity and affinity, batch after batch.
• Integrated reading solutions: Mabtech IRIS™ 2 with the patented RAWspot™ algorithm provides the highest precision in spot detection, counting, and polyfunctional assessment available.

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Applications

T cell applications

ELISpot and FluoroSpot is commonly used to investigate antigen-specific immune responses and to discriminate between subsets of activated T cells. This is applied in studies of infectious diseases, cancer, allergies, and autoimmune diseases.

Immunotherapy

Both ELISpot and FluoroSpot are powerful potency assays in CAR-T, TIL, and gene therapy development, giving clear insight into cytokine release such as IFN-γ, Granzyme B, and TNF-α from engineered T cells or virally transduced cells. These assays align with FDA recommendations for immunotherapy potency testing.

Infectious disease

ELISpot has been essential for mapping cellular immune responses in infectious disease research ranging from TB and SARS-CoV-2 to malaria and HIV, by quantifying pathogen-specific cytokine-secreting cells. The assay’s sensitivity and robustness at the single-cell level make it a key tool for both diagnostic and vaccine-driven investigations.

Vaccine Research

In vaccine research, IFN-γ ELISpot is the gold standard to define vaccine efficacy by measuring the capacity to elicit potent T cell responses. IFN-γ/IL-2/TNF-α FluoroSpot is heavily used to define vaccine efficacy by measuring the capacity to elicit potent T cell responses. IFN-γ/IL-2/Granzyme B FluoroSpot is another great combination to assess both Th1 and cytotoxic T cells in the same well, saving on precious samples.

Diagnostic assays based on ELISpot are also readily available, including tests to detect patients with tuberculosis or SARS-CoV-2 infection by measuring IFN-γ secretion from T cells responding to defined antigens. And please note: ELISpot is not limited to IFN-γ. Mabtech offers ELISpot in over 40 different cytokines and other analytes to answer all your research questions.

Mabtech's ELISpot Assays are available for the full range of cytokines:

B cell applications

B cell ELISpot and B cell FluoroSpot are two of few assays that can measure immunoglobulins directly upon secretion. One can assess total immunoglobulin secretion by antibody-secreting cells (ASCs) or memory B cells, as well as antigen-specific B cell responses from both cell types using the antigen of interest as a capture or detection reagent.

Detect acute or memory B cell responses

B cell ELISpot can be used to answer several research questions in infection and vaccine research. One can assess early acute ASC responses following vaccination or during infection by isolating the cells directly from donor blood and using them in the assay. Additionally, researchers can assess the memory B cell response elicited by a vaccine or following infection to assess the longevity of the B cell response. Here, isolated PBMCs are stimulated with polyclonal activators to induce memory B cells to secrete antibodies for assessment in the assay.

Evaluate cross-reactive B cells

B cell FluoroSpot can be used to answer several research questions in infection and vaccine research. One can assess multiple immunoglobulin isotypes, subclasses, and even antigens! By using reverse B cell FluoroSpot, multiple antigens can be used to detect single and cross-reactive B cells. This adds new, added layers to your B cell research and can help guide new vaccine development and assessment.

ELISpot assays for human Immunoglobulins:

Other Species also available.

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18.05.2026