The gold standard for immune monitoring
The ELISpot assay is a robust and straightforward tool for detecting and quantifying antigen-specific immune responses at the single-cell level. For almost 40 years, researchers have trusted Mabtech's ELISpot kits for their reliability and high sensitivity in capturing both T cell and B cell responses.
So how does ELISpot work? The assay works similarly to ELISA, but instead of measuring an analyte in solution, it detects the cells secreting that analyte (see image). Immune cells are stimulated in wells pre-coated with capture antibodies specific to the analyte of interest. Responding cells secrete the analyte, which is immediately captured by the antibodies on the plate. After incubation, the cells are washed away, and detection antibodies are added, followed by visualization reagents to reveal spots. Each spot represents an individual responding cell, offering a precise and quantifiable measure of immune activity.
ELISpot vs ELISA
What’s the difference between ELISA and ELISpot? While both are sensitive sandwich assays, ELISA measures the analyte concentration in a solution, like plasma or serum, whereas ELISpot detects it directly from cells as they secrete it. ELISA provides analyte concentration, while ELISpot quantifies the number of secreting cells (spot-forming units, SFU) and their secretion levels (relative spot volume, RSV). This makes ELISpot especially sensitive, as it captures the analyte continuously throughout the cell stimulation process.
Additionally, as ELISpot captures target proteins immediately after secretion, the method can detect cytokines that otherwise are too diluted or disappear from samples. For example, ELISpot can detect analytes that are rapidly degraded by proteases (IL-2), are quickly taken up by bystander cells (IL-4), or bind to soluble receptors (TNF-α).
ELISpot was instrumental in the development of the COVID-19 vaccines and in characterizing immune responses following infection. Mabtech's ELISpot kits played a pivotal role in much of this research.
Why Mabtech ELISpot?
Mabtech's ELISpot kits are trusted by clinical trial labs, pharmaceutical companies, and CROs for their robustness and superior sensitivity in detecting antigen-specific immune responses at the single-cell level. Recognized as the gold standard for evaluating antigen-specific T cells and B cells, ELISpot is ideal for tracking immune responses throughout pre-clinical studies and clinical trials. Whether you're assessing vaccine efficacy, assessing immune modulation, or monitoring cell therapy potency, Mabtech's ELISpot kits seamlessly integrate into immune monitoring workflows, providing the high-quality, reliable data critical for advancing therapeutic success.
"We have used Mabtech ELISpot antibodies and reagents for several years. The high quality and standard and competitive price will keep us hooked for years to come."
Cecilia Lindestam Arlehamn
Division of Vaccine Discovery
La Jolla Institute for Immunology (LJI), USA
"Our research group uses Mabtech products for a variety of applications, especially investigating immunological responses in wildlife. We have been able to develop new assays for detecting diseases."
Michele Miller
Faculty of Medicine and Health Sciences
Stellenbosch University, South Africa
T cell profiling
ELISpot is commonly used to investigate antigen-specific immune responses and to discriminate between subsets of activated T cells. It's often utilized in researching infectious diseases, cancer, allergies, and autoimmune diseases. In vaccine research, ELISpot is the gold standard to define vaccine efficacy by measuring the capacity to elicit T cell responses, for example by assessing IFN-γ secretion. Diagnostic assays based on ELISpot are available, including tests to detect patients with tuberculosis or SARS-CoV-2 infection by measuring IFN-γ secretion from T cells responding to defined peptides.
Human IFN-γ ELISpot
IFN-γ secretion by peripheral blood mononuclear cells (PBMCs) incubated overnight without stimuli or with anti-CD3, phytohaemagglutinin (PHA) , or purified protein derivative (PPD).
Human IgG and IgA ELISpot
PBMCs were collected before and after vaccination against influenza and secretion of influenza A (H1N1)-specific IgG and IgA was analyzed by ELISpot.
B cell profiling
The B cell ELISpot assay is one of few assays measuring immunoglobulins directly upon secretion. There are two strategies: First, B cell ELISpot can be used to assess antibody-secreting cells (ASCs). Due to its sensitivity, the method enables the identification of rare ASCs to a specific antigen. Secondly, you can evaluate circulating antigen-specific memory B cells after polyclonal activation. B cell ELISpot is thus regularly used to detect B cell responses elicited by infection or vaccination. Read more about B cell ELISpot here!
ELISpot kit formats
At Mabtech, we offer a range of ELISpot products in different kit formats and with different detection reagents. From super flexible to ready-to-go kits, we've got you covered. Find the kit format that fits your workflow below. And while we prefer the ALP detection system, we also offer our kits with HRP detection reagents.
Our most flexible ELISpot kit option available when you already have plates and ELISpot substrate.
Total assay time:
- 3-day protocol
- 2-step detection
Kit size options:
- Reagents for 4 plates
- Bulk options available upon request
Our most popular ELISpot kits with everything you need for the assay including a positive control (for human and NHP kits).
Total assay time:
- 2-day protocol
- 2-step detection
Kit size options:
- 2- or 10-plate kits
- 100-plate kits for certain analytes
- Bulk options available upon request
The benefits of the Plus format, but with a one-step detection system. Ideal when time is of the essence!
Total assay time:
- 2-day protocol
- 1-step detection
Kit size options:
- 2- or 10-plate kits
- 100-plate kits for certain analytes
- Bulk options available upon request
A complete ELISpot kit to answer your specific scientific question with an included PepPool and positive control. Just add cells!
Total assay time:
- 2-day protocol
- 1- or 2-step detection
Kit size options:
- 1-plate kit
- Bulk options available upon request
Mabtech ASTOR 2 – the next generation ELISpot reader
Unlock the full potential of your ELISpot assays with Mabtech ASTOR 2. Its unmatched analytical precision, combined with user-friendly software, transforms complex data into clear and tangible results, making it the ELISpot tool for cutting-edge research.
Discover why ASTOR 2 sets the standard for ELISpot readers in accuracy and simplicity. Connect with our team for a demo and see how ASTOR 2 can transform your research or be implemented into automated setups.
At Mabtech, we aim for awesome every time
Find antigen-specific cells
With ELISpot, you can detect any cell secreting the analyte of interest. The assay is considered the gold standard for detecting and enumerating antigen-specific T cells and B cells. We've made it easy to design your ELISpot setup for success. Learn more in our Knowledge Hub or download our step-by-step ELISpot guide below.
User-friendly software
Building on 35+ years of ELISpot technology, we created a reader to meet our high standards. Introducing the next-generation ELISpot reader ASTOR 2, powered by the innovative RAWSpot™️ technology, designed to deliver unmatched accuracy and sensitivity in spot detection. We paired our novel detection algorithm with the most intuitive software available. Simply load your plate, hit "Read," and get your results. With ASTOR 2, it's not just about reading spots – it's about pioneering a new era of ELISpot analysis.
We're here to help
At Mabtech, our mission is to aid researchers in reaching their goals. We do this not only by providing the best possible monoclonal antibodies, immunoassay kits, and instruments but also through webinars, educational blogs, workshops, and attending world-renowned immunology meetings. Check out our entire Knowledge Hub for inspiration, troubleshooting, guides, and more!
