Mouse 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

To determine the level of secreted adiponectin in control or SEPT1-knockdown 3T3-L1 adipocytes, cells were washed with PBS, and 500 µl of serum-free IMDM containing 100 nM insulin and 1% penicillin/streptomycin were added per well in a 12-well plate. 3T3-L1 adipocytes were incubated for 24 h at 37°C. Then the medium was recovered and the remaining material was centrifuged for 5 min at 500 g at room temperature to remove cell debris. Adiponectin levels in the collected media were measured with an enzyme-linked immunosorbent assay (ELISA) (DY1119, R&D Systems GmbH) following the manufacturer's instructions. Cells were washed with PBS and lysed to determine protein concentration. For quantification, adiponectin levels were normalized to the respective protein concentration.

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Manufacturer protocol

Download the product protocol from R&D Systems for Mouse Adiponectin/Acrp30 DuoSet ELISA below.

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