Human Adiponectin/Acrp30 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

Adiponectin assay

Blood samples were collected in the morning after overnight fasting. The serum samples were clotted and centrifuged at 2000×g for 10 min and immediately frozen at -80°C for further analysis. Adipocytokine levels were measured using an enzyme-linked immunosorbent assay (ELISA) kit (R&D Systems, MN, USA), and adiponectin levels (with the sample diluted 4,000-fold) were measured using the DuoSet® ELISA kit (DY1065; R&D Systems). The analytical methodology and technical procedures were performed according to the manufacturer’s protocol.

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Manufacturer protocol

Download the product protocol from R&D system, Minneapolis, MN, USA for Human Adiponectin/Acrp30 DuoSet ELISA below.

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