Mouse Prolactin 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

Mice were housed individually for 1 week before the experiments were started and kept single-caged throughout the experiments in temperature-, humidity-, and light (12 h light: 12 h darkness cycles)-controlled rooms. Samples for hormone determinations were obtained by tail bleeding between 08.00 and 10.00 a.m., after 8 and 15 dpi, as well as from uninfected mice. Blood was centrifuged during 10 min at 1,500 rpm at 25°C and serum was stored frozen at −20°C until analyzed. Corticosterone levels were determined by RIA (MP Biomedicals, New York, USA) as previously described [41], whereas PRL concentration was evaluated by specific ELISA kit, according to the manufacturer's specifications (R&D Systems, Minneapolis, USA).

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Manufacturer protocol

Download the product protocol from R&D Systems for Mouse Prolactin DuoSet ELISA below.

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