Rat JE/MCP-1/CCL2 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

After the last dosage, rats went through fasting for 12 hours before blood was collected from abdominal artery. Collected blood was centrifuged at 3000 rpm for 10 minutes and the serum was separated. Glucose, insulin, total cholesterol (TC), total triglyceride (TG), tumor necrotic factor a (TNF-a), interleukin-6 (IL-6), intracellular adhesion molecule 1 (ICAM-1), vascular-cell adhesion molecule 1 (VCAM-1) and monocyte chemoattractant protein-1 (MCP-1) levels were then determined using corresponding enzyme-linked immunosorbent assay (ELISA) kits (R&D Systems, USA).

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Manufacturer protocol

Download the product protocol from R&D Systems for Rat JE/MCP-1/CCL2 DuoSet ELISA below.

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