THUNDER™ Human GM-CSF TR-FRET Biomarker Assay Kit

CSF2|Granulocyte-macrophage Colony-stimulating Factor

The THUNDER™ human GM-CSF assay kit is a homogeneous time-resolved Förster resonance energy transfer (TR-FRET) sandwich immunoassay. The assay workflow consists of a one-step reagent addition. Human GM-CSF in the cell supernatant sample is detected with a pair of fluorophore-labeled antibodies in a simple "add-incubate-measure" format (no wash steps). One antibody is labeled with a donor fluorophore (Europium chelate; Eu-Ab1) and the second with a far-red acceptor fluorophore (FR-Ab2). The binding of the two labeled antibodies to distinct epitopes on the target cytokine takes place in solution and brings the two dyes into close proximity. Excitation of the donor Europium chelate molecules with a flash lamp (320 or 340 nm) or a laser (337 nm) triggers a FRET from the donor to the acceptor molecules, which in turn emit a TR-FRET signal at 665 nm. Residual energy from the Eu chelate generates light at 615 nm. The signal at 665 nm is proportional to the concentration of human GM-CSF in the cell supernatant. Data can be expressed as either the signal at 665 nm or the 665 nm/615 nm ratio. A distinct advantage of the THUNDER™ TR FRET technology is that it does not require addition of potassium fluoride to ensure signal stability. The non-toxic nature of THUNDER™ makes it an ideal candidate for multiplexing with other assay technologies using a sequential assay protocol.

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