Human GM-CSF DuoSet ELISA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Bring all reagents to room temperature before use. Allow all components to sit for a minimum of 15 minutes with gentle agitation after initial reconstitution. -Working dilutions should be prepared and used immediately.	-A standard curve should be generated for each set of samples assayed. -It is suggested to start Reagent Diluent optimization for serum and plasma samples by using PBS supplemented with 10-50% animal serum. -Do not use buffers with animal serum to reconstitute or dilute the Detection Antibody or Streptavidin-HRP B.

Upstream tips
-Bring all reagents to room temperature before use. Allow all components to sit for a minimum of 15 minutes with gentle agitation after initial reconstitution. -Working dilutions should be prepared and used immediately.
Protocol tips
-A standard curve should be generated for each set of samples assayed. -It is suggested to start Reagent Diluent optimization for serum and plasma samples by using PBS supplemented with 10-50% animal serum. -Do not use buffers with animal serum to reconstitute or dilute the Detection Antibody or Streptavidin-HRP B.

Publication protocol

MGIA supernatants were analysed to assess cytokine concentrations by enzyme-linked immunosorbent assay (ELISA). The levels of following cytokines were measured: interferon-gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), interleukin (IL)−12p40, IL-10, IL-17, IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon-gamma-induced protein 10 (IP-10). The concentrations of IFN-γ, IL-12p40 and IL-6 were measured using BD OptiEIA kits (Becton Dickinson, UK), while TNF-α, GM-CSF and IP-10 DuoSet ELISA kits were obtained from R&D Systems (Abingdon, UK), and IL-10 ELISA MAX Standard and IL-17 ELISA MAX Deluxe from BioLegend (London, UK). Assays were performed as described by the manufacturers.

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Manufacturer protocol

Download the product protocol from R&D Systems for Human GM-CSF DuoSet ELISA below.

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