Cytokine ELISPOT Assay

All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

Cytokine ELISPOT assay, a powerful experimental technique, can be used to measure cytokine secreting cells and assess the effects of immune-modulating therapies in oncology and immunology fields. With unrivaled expertise and experience in various immunotherapy discoveries and development, Creative Biolabs is confident in providing optimized cytokine ELISPOT assay service to best suit your program requirements.

As a Highly Sensitive, Quantitative Assay to Measure Cytokine Secreting Cells

As a highly sensitive, quantitative Assay, ELISPOT assays are easier to detect a much smaller amount of cytokine and chemokine produced by individual immune cells. ELISPOT assays use high-affinity antibodies against different epitopes of a single cytokine/chemokine.

Compared with ELISA, the ELISPOT assay can not only detect cytokine response at the single-cell level but also measure specific cytokine-producing cells with high sensitivity, even if the cytokine secreted into the supernatant is lower than the level detected by ELISA.

The Foundation of Cytokine ELISPOT Assay

Cytokine ELISPOT is based on the following principle: cytokines secreted by immune cells are captured by antibodies on a matrix, and a secondary detection antibody combines with a substrate to precipitate at the location of the captured cytokines to form visible spots.

Cytokine ELISPOT Assay

The basic ELISPOT protocol is designed to quantify the number of cells secreting specific cytokine/chemokine. The critical first step involves binding the high-affinity anti-cytokine Ab to a solid matrix. When the cells of interest are incubated in the plate, they secrete the cytokine/chemokine of interest. Then, the interest secreted protein binds to the pre-captured Abs immediately. This binding protein is recognized by a secondary enzyme-linked Ab. A colorimetric substrate is used to produce a dark precipitate or "spot" that marks the location of the protein-producing cell. The resultant spots are permanent and can be quantified visually, microscopically, or electronically.

Cytokine ELISPOT Applications

Detecting cytokine secreted
High throughput epitope mapping
Measuring and assessing the immune response of immune cells
Monitoring immunogenicity of vaccines, cell therapies, and immune-modulating therapies (CAR-T, checkpoint inhibitors, cytokines, agonists, and adjuvants)

Creative Biolabs will be with you at every step of your way. For more details of our Cytokine ELISPOT assay service, please reach out to our scientists to learn how we can be involved in your project.

References

Ji, Niannian, and Thomas G. Forsthuber. "ELISPOT techniques." Multiple Sclerosis. Humana Press, New York, NY, 2014. 63-71.
Klinman DM, Nutman TB. ELISPOT assay to detect cytokine‐secreting murine and human cells. Current Protocols in Immunology. 1994 Jun;10(1): 6-19.
Tang Y, et al. The co-stimulation of anti-CD28 and IL-2 enhances the sensitivity of ELISPOT assays for the detection of neoantigen-specific T cells in PBMC. Journal of Immunological Methods. 2020 Sep 1; 484: 112831.

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