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NCL Method ITA-27

By Edward Cedrone, Timothy Potter, Barry Neun, Abby Tyler, Marina Dobrovolskaia

Multiplex Enzyme-Linked Immunosorbent Assay (ELISA) for Detection of Human Cytokines in Culture Supernatants

Listed in Datasets | publication by group NCL Protocols

Version 1.0 - published on 11 Jun 2021 doi:10.17917/D4XK-WH24 - cite this


Cytokines, chemokines and interferons (IFN) are soluble mediators of inflammation and a variety of responses that orchestrate both innate and adaptive immunity in health and disease [1-10]. Understanding the induction of these biomarkers in response to a drug product helps to establish the product's safety profile and shed light on the mechanism of action or mechanism to toxicity. This document describes experimental procedure for analysis of culture supernatants by multiplex ELISA to detect presence of type I IFNs (IFNa, a, IFNb, b, IFNw), type II IFN ( IFN-g), type III IFN ( IFN-l), cytokines and chemokines (IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p70, IL-13, IL-15, IL-17, IL-21, IL-22, IL-23, IL-27, IP-10, MCP-1, MCP-2, MIP-1α, MIP-1β, RANTES, TNF- α) in culture supernatants. NCL protocol ITA-10 should be referred to for details of preparation of culture supernatants. Protocol for single-plex ELISA (ITA-22, ITA-23, ITA-24 and ITA-25) can also be considered. The advantage of this protocol over single-plex ELISA is that it allows simultaneous analysis of a broad spectrum of markers.

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