How does an ELISpot assay work?

The enzyme-linked immunospot (ELISpot) assay is a highly sensitive immunoassay that measures the frequency of cytokine-secreting cells at the single-cell level. In this assay, cells are cultured on a surface coated with a specific capture antibody in the presence or absence of stimuli.

How do you dry ELISpot plates?

Both polystyrene-bottomed and PVDF membrane-bottomed plates should be dried without a lid upside down at room temperature. When dry, PVDF membrane-bottomed plates with enzymatically colored spots should be stored in the dark at a dry place at room temperature to prevent bleaching of spots.

What is ELISpot assay used for?

ELISpot assays are quantitative and measure key cellular functions of immune system cells. ELISpot has been used to assess both adaptive and innate immune responses.

What types of cells can be used in ELISpot?

With the ELISpot, you can study antigen-specific cytokine responses, antibody specific secreting cells, tumor antigens, granzyme B and Perforin release by T cells, vaccine efficacy, epitope mapping, cytotoxic T-cell activity, detection of IL-4, IL-5, and IL-13, vaccine-induced antibody responses, antigen-specific …

Which kind of antibodies are used in EliSpot assay?

ELISpot assays employ the sandwich enzyme-linked immunosorbent assay (ELISA) technique. Either a monoclonal or polyclonal antibody specific for the chosen analyte is pre-coated onto a PVDF (polyvinylidene difluoride)-backed microplate.

How do you do an EliSpot?

ELISPOT is performed using a PVDF or nitrocellulose membrane 96-well plate pre-coated with an antibody specific to the secreted protein. Cells are added to the plate and attach to the coated membrane. Cells are then stimulated and the secreted protein binds to the antibody.

Is ELISPOT more sensitive than ELISA?

The level of detection by ELISPOT was 10 to 200 times more sensitive than ELISA performed on culture supernatants. We also demonstrated that the solubilized ELISPOT can detect cytokine secretion by cells with greater sensitivity than conventional ELISA.

What is IFN gamma ELISPOT?

Interferon-gamma (IFN-γ) is often used as a cytokine endpoint in ELISpot assays and has a wide range of biological functions in the context of an immune response, including the activation of macrophages, up-regulation of major histocompatibility complex I and II, and induction of IgG production from activated B cells.

How do you do an ELISPOT?

What is ImmunoSpot assay?

At the unprecedented resolution of up to 1 in 800,000, ImmunoSpot® assays measure effector molecule secretion by individual T cells that have been stimulated by an antigen. ImmunoSpot® assays therefore provide information on the numbers of the antigen-specific T cells present in a test cell population, typically PBMC.

What is Elisa sandwich?

A sandwich ELISA measures antigen between two layers of antibodies (capture and detection antibody). The target antigen must contain at least two antigenic sites capable of binding to antibodies. Monoclonal or polyclonal antibodies can be used as the capture and detection antibodies in sandwich ELISA systems.

How is the ELISpot assay used by mabtech?

Mabtech has worked extensively for more than 20 to 30 years to optimize the ELISpot protocol. New proteins are continuously added to the range of analytes that can be analyzed by ELISpot. The ELISpot assay is carried out in a 96-well plate, and an automated ELISpot reader is used for analysis.

What kind of antibodies do we use at mabtech?

Mabtech supplies a wide range of antibodies to cytokines, immunoglobulins and apolipoproteins. We are specialized in ELISpot, FluoroSpot and ELISA and provide reagents for use in many research fields.

What is the limit of detection for ELISpot?

The limit of detection typically achieved can be 1 in 100,000 cells. The high sensitivity of the assay makes it particularly useful for studies of the small population of cells found in specific immune responses. Mabtech has worked extensively for more than 20 to 30 years to optimize the ELISpot protocol.

What are different types of ELISpot research kits?

We offer ELISpot kits in different formats for different research needs: from flexible kits that include only the core reagents to complete pre-coated kits made for studying immune response to specific pathogens.

How does an ELISpot assay work? The enzyme-linked immunospot (ELISpot) assay is a highly sensitive immunoassay that measures the frequency of cytokine-secreting cells at the single-cell level. In this assay, cells are cultured on a surface coated with a specific capture antibody in the presence or absence of stimuli. How do you dry ELISpot plates?…