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ECIA™ Intracellular Cytokine Staining Assay

Cytokines are intercellular signal transduction molecules that closely mediate and adjust immune response, inflammatory reaction, promotion of hematopoietic cell differentiation and restoration of damaged tissues. Cytokines are involved in many different yet important pathways including the induction of T cell proliferation by IL-2, the induction of anti-viral protein expression by IFN gamma, and the inhibition of viral gene expression and replication by TNF alpha. The dysregulation of cytokine expression has been widely associated with many diseases and thus, the detection of cytokine expression levels is of vital importance for diagnosis as well as therapy development purposes.

The quantitative and activity detections of cytokine levels from a population of cells have been unable to provide enough information in some occasions. Therefore, a novel technology has been developed to meet these specific demands. Intracellular cytokine staining (ICS) assay coupled with flow cytometry can be utilized to detect the antigen-specific T cell responses at a single cell level for both clinical studies and scientific researches. This technology harnesses the particular advantage of enabling the simultaneous assessment of multiple phenotypic, differentiation and functional parameters pertaining to responding T cells, most notably, the expression of multiple effector cytokines.

Creative Biolabs has been providing ECIA™ intracellular cytokine staining services for customers all over the world for many years. With our extensive experience and expertise, your assays will be designed and conducted in the most efficient and cost-effective way. We realize that there are differences in various projects and objectives, therefore we frame our services in a highly-customized fashion by communicating with customers thoroughly for better solutions and optimization. We are willing to hear questions and suggestions from customers to make sure that the results of our services contribute to your project in a broader perspective.

ICS is achieved through blocking the secretion of protein mediated by Golgi apparatus by utilizing in vitro polyclonal activators or specific antigen-stimulated cells and cytokine transmembrane secretion blockers (e.g. brefeldin A), allowing the accumulation of cytokines in the cytoplasm. Fluorescence-labeled antibodies are then used to detect the cytokines. ICS can reflect characteristics of a single cell and display the specificity of cell groups, which directly indicates the in vivo cell state and the level of protein in the cytoplasm. After antibody staining, flow cytometry is used for analysis of the quantity and quality of relevant cells.

General Workflow of ECIA™ Intracellular Cytokine Staining Assay


Featured Applications of ECIA™ Intracellular Cytokine Staining Assay

The Protocol of Intracellular Cytokine Staining Fig.1 The Protocol of Intracellular Cytokine Staining.

Reference:

  1. Kenneth D, et al. “Single-cell phospho-specific flow cytometric analysis demonstrates biochemical and functional heterogeneity in human hematopoietic stem and progenitor compartments.” Blood 117 (2011): 4226-4233.



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