ELISPOT

IFN-γ ELISPOT,
the golden standard
for immune monitoring of
antigen-specific T cell responses

Enumeration of antigen-specific T cells using the enzyme-linked spot forming assay, better known as ELISPOT, is widely applied in both commercial and academic studies. The familiarity, ease of use and immediate understanding of data all favor this assay for tracking antigen-specificity in animal and human studies. Even though ELISPOT protocols are established and run on a frequent basis in multiple labs, however, both the intra-assay and inter-laboratory variance for ELISPOT is known to be challenging, stressing the importance of letting experts taking care of such studies. Such variability has been shown to come from both the laboratory work and from the analysis part. We have the necessary expertise in both in-house and deliver accurate data with a rapid turnaround, freeing up time and energy for you to carry out other activities.

ImmuMap have established ELISPOT assays for detection of IFN-γ and TNF-α, and we offer developing also assays for detection of other cytokines such as IL-2, granzyme B and others. We have experience in using a wide range of stimulants such as non-specific agents, compounds from our clients, short or long peptides or whole proteins. Further, we offer taking care of each of the laboratory and data analysis parts of the ELISPOT in addition to performing the full analysis if you prefer this. We understand your need for confidentiality and if needed, we can perform the analysis blinded with no information about the stimulants or peptides used. In our experience, however, the more information we obtain prior to running the analyses, the better we can customize the assay and subsequently obtain better data.

No co-detection in addition to the cytokine in question is possible so ELISPOT does not provide information about e.g. whether the responding cells are CD4 or CD8 T cells. Normally we measure the cytokine secretion from the heterogenous PBMC fraction, however, we also offer including a pre-ELISPOT separation step for testing e.g. the CD8 cells only.

Applications of ELISPOT:

  • Detection and enumeration of functional antigen-specific T cells or other cells responding to the stimulant in question in a single sample in in vitro studies.
  • Immune monitoring of pre-clinical or clinical in vivo samples using the method preferred by the regulatory authorities for such studies
  • Confirmation and further characterization of cytokine secretion by antigen-specific T cells or other cell subsets as a follow up on previous intracellular cytokine staining data.

We understand the need for linking together data obtained using ELISPOT with flow cytometry-based assays focused on antigen-specificity such as MHC multimer staining and intracellular cytokine staining, and therefore offer analyzing customer samples with either one, two or all three assays in parallel, see Intra-cellular cytokine staining and MHC multimer staining for further information about these methods

Contact our staff today for a discussion on how we can help you.

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ImmuMap - DTU Science Park, Diplomvej 377, 2800 Kgs. Lyngby, Denmark - Tel: +45 6178 1882

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