Intracellular Cytokine Staining Service

ProImmune offers rapid and reliable flow cytometric detection of human IFN-gamma, IL-2 and TNF-alpha cytokine production in CD4+ and CD8+ T cells through its experienced applications team.

By outsourcing your intracellular cytokine staining experiments to ProImmune, you take advantage of our technical proficiency and save time by passing collection and analysis of data to our experts.

Shipping Samples

Application Notes

Sample Protocol

Request Quote

Intracellular cytokine staining by flow cytometry is a powerful technique that allows the analysis of individual cells in a mixed population. The procedure relies upon the stimulation of T cells in the presence of an inhibitor of protein transport, in order to retain the cytokines inside the cell. Cells are first stimulated with antigen, followed by staining with antibodies specific for extracellular epitopes, such as CD4 and/or CD8. Membrane permeabilization and intracellular cytokine staining follow. The frequency of cells that produce a particular cytokine is measured using fluorescent antibodies.

Intracellular cytokine staining can be used as an immune monitoring tool to measure the immune response to known antigens, or for epitope discovery, combined with ProImmune’s REVEAL & ProVE® technology, to identify and/or validate novel T cell epitopes. Compared to the ELISPOT technique, flow cytometric detection of intracellular cytokines allows for simultaneous phenotyping and gating of cells, to select the specific live lymphocyte population.

Multi-center clinical trials pose the challenge of collecting, shipping and processing samples in a way that ensures consistency and repeatability. Sample logistics and preparation is probably the most important step in setting up successful assays for monitoring cell mediated immune responses. We have the experience and the capability to collect your samples wherever you are in North America and Europe and process them for maximum uniformity; sample preparation and shipping.

Our intracellular cytokine staining service is based on a standardized format, which follows this step-by-step process:

(1) Customer cryopreserves cells in accordance with optimized protocol recommended by ProImmune or optimized customer protocol;

(2) ProImmune collects cells from customer site using validated shipper (customer may also use their own shipper),

a. ProImmune carries out HLA tissue typing as an option if required,
b. ProImmune carries out custom peptide synthesis as an option if required;

(3) ProImmune thaws cells with optimized protocol and carries out intracellular cytokine staining in accordance with standardized procedure.

(4) Customer receives assay report, including flow cytometry staining data, plus raw data .fcs files on CD.

The delivery time for a project will vary depending on the specific requirements, but a turnaround time of 3-5 weeks (including peptide synthesis and shipment of cells) can be expected for an average project.

Contact us if you require intracellular staining for other cytokines.

Example staining for an intracellular cytokine staining experiment.

Request the Cellular Analysis Services Survey Form to obtain a quotation for your precise requirements.

Co-staining for MHC Pentamers and intracellular cytokines

Overview of Cellular Analysis Services

Cellular Analysis Services (PDF)