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ELISpot kits

Visualize the secretory products of individually activated or responding cells with our high-sensitivity ELISpot assays. Part of a portfolio of over 4k ELISA kits and immunoassays with 15k+ citations, these tools enable precise monitoring of immune responses at the single-cell level for unmatched experimental clarity.
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What is an ELISpot assay?

ELISpot (Enzyme-Linked ImmunoSpot) is a sensitive immunological assay closely related to the ELISA technique that was developed to detect and quantify secreted proteins, such as cytokines, growth factors, or antibodies, at the single-cell level.

ELISpot assays allow visualization of the secretory product of individually activated or responding cells. Each spot that develops in the assay represents a single reactive cell. They provide valuable insights into the immune response at the single-cell level and are essential in immunology and vaccine research.

We offer a range of ready-to-use T-cell ELISpot kits as well as ELISpot kits with uncoated plates for more flexibility.

How ELISpot assays work

The ELISpot assay is based on the sandwich ELISA technique but adapted to measure secretion at the single-cell level:

  1. A PVDF membrane is coated with a capture antibody specific to the target analyte (eg cytokine or immunoglobulin)
  2. Cells are added and cultured under stimulating or resting conditions
  3. Secreted analytes are captured immediately by the immobilized capture antibody
  4. A biotinylated detection antibody binds to the captured analyte
  5. An enzyme conjugate (eg streptavidin-alkaline phosphatase) is added
  6. Substrate produces an insoluble precipitate at the secretion site
  7. Each spot corresponds to a single secreting cell

This enables direct quantification of antigen-specific immune responses.

How ELISpot results are read

ELISpot results can be analyzed using:

Results are typically expressed as:

Spot size and intensity can also provide qualitative insights into secretion strength.

ELISpot protocol overview

A typical ELISpot workflow includes:

Check our detailed ELISpot protocol guide

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ELISpot vs ELISA - key differences

Feature
ELISpot
ELISA
Detection level
Single-cell
Bulk sample
Sensitivity
Very high
High
Output
Frequency of secreting cells
Total protein concentration
Use case
Immune response profiling
Protein quantification
Applications
T-cell assays, vaccine studies
General protein assays

ELISpot is ideal for detecting rare antigen-specific cells, while ELISA is used for quantifying total protein levels in a sample.

What research applications are ELISpot used in?

T-cell ELISpot assays are mainly used to measure antigen-specific T-cell responses in a variety of applications:

Vaccine research and development

Quantify how well candidate vaccines stimulate antigen‑specific T cells. Provide sensitive readouts of protective cellular immunity.

Infectious diseases studies

Detect T-cell responses to pathogens such as viruses (HIV, influenza, SARSCoV2), bacteria (tuberculosis), and parasites.

Cancer immunology and immunotherapy

Measure tumor-specific T-cell activity. Evaluate patient responses to checkpoint inhibitors, CART therapies, and cancer vaccines.

Basic immunology studies

Investigate cytokine secretion profiles (IFNγ, IL-2, IL-10, TNFα) from individual T cells. Enables frequency analysis of rare antigen-specific populations that are difficult to detect with other methods.

Why use ELISpot assays?

ELISpot assays provide an accurate and sensitive method to assess the T-cell responses. Their key benefits are:

Our T‑cell ELISpot assays provide sensitive, single‑cell resolution for key cytokines such as IFN Gamma, IL‑2, IL‑4, and IL‑10, enabling precise immune monitoring across research and clinical applications.

ELISpot assays are part of a broader immunology workflow. You may also require:

Explore more ELISpot resources

To support your workflow, explore:

FAQs

What is the main advantage of ELISpot over other immunoassays?

ELISpot enables detection at the single-cell level, making it more sensitive than bulk assays like ELISA.

What types of cells can be used in ELISpot assays?

Commonly used cells include PBMCs, T-cells, B cells, and other immune cell populations.

Are ELISpot assays quantitative?

Yes, results are expressed as spot counts, reflecting the frequency of secreting cells.

Can ELISpot detect multiple cytokines?

Yes, multiplex ELISpot assays can detect multiple analytes simultaneously, depending on assay design.

Do I need specialized equipment?

Automated ELISpot readers improve accuracy and reproducibility, but manual counting is also possible.

References

  1. Czerkinsky, C. C., Nilsson, L.-Å., Nygren, H., Ouchterlony, Ö. & Tarkowski, A. Enzyme-linked immunospot assay for the detection of antigen-specific antibody-secreting cells. J. Immunol. Methods
  2. Moodie, Z., Price, L., Gouttefangeas, C. & Janetzki, S. Response determination criteria for ELISpot assays revisited. Cancer Immunol. Immunother.
  3. Janetzki, S. et al. Guidelines for the automated evaluation of ELISpot assays. Nat. Protoc.
  4. Schmittel, A., Keilholz, U. & Scheibenbogen, C. Evaluation of the interferon-γ ELISpot assay for immunotherapy monitoring. J. Immunol. Methods
  5. Lehmann, P. V., Zhang, W. & Unique strengths of ELISpot for T cell assays. Cell. Mol. Immunol.
  6. Cox, J. H., Ferrari, G. & Janetzki, S. Measurement of cytokine release at the single-cell level using ELISpot. Methods
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