Human Total HO-1/HMOX1 DuoSet IC 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

Homogenized tissues at 100 mg ml-1 in 1 × phosphate buffered saline+0.5% Triton X-100+protease inhibitor cocktail (Roche, Indianapolis, IN, USA) were centrifuged at 4 °C at 4000 rpm for 5 min. Supernatant was collected and stored at −80 °C. DuoSet ELISA kit determined GDNF concentration (DY212, R&D systems) according to the manufacturer's instructions. Samples were run with biological (n>4 per group) and technical (n=3) replicates.

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Manufacturer protocol

Download the product protocol from R&D Systems for Human Total HO-1/HMOX1 DuoSet IC ELISA below.

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