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Simultaneous detection of IL-6 and IL-8 secretion by cell lines using AlphaLISA and AlphaPlex 545 reagents


Cells react to various stimuli in the body by secreting modulator proteins called cytokines. These proteins bind to specific receptors to generate a response from the targeted cell. These responses range from cell growth, mobility, to alterations in differentiation and function, and even cell death. Cytokines are involved in many pathological pathways, including inflammation and cancer. As such, they are as such interesting research targets.
Immunoassays are the primary method used to measure production and modulation of cytokines by cells. However, the majority of these technologies are work-intensive, require large amounts of sample and can only analyze one cytokine per assay.
A new Alpha technology has been developed to allow the analysis of two different cytokines in the same sample. The combination of beads based on AlphaLISA® (europium) and AlphaPlex™ 545 (terbium) chemiluminescence allows for a fast, homogeneous assay using as little as 5μL of cell supernatant. Here, we demonstrate the value of this assay on the detection of both IL-6 and 8, key cancer biomarkers, from two cell lines stimulated by IL-1b.
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