Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Store the unopened kit at 2-8 °C. -Bring all reagents to room temperature before use. -Reconstitute the Human Cytochrome c Standard with deionized or distilled water. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions.	-When mixing or reconstituting protein solutions, always avoid foaming. -Substrate Solution should remain colorless until added to the plate. -Keep Substrate solution protected from light. -Stop Solution should be added to the plate in the same order as the Substrate Solution. -Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the Substrate Solution.

Upstream tips
-Store the unopened kit at 2-8 °C. -Bring all reagents to room temperature before use. -Reconstitute the Human Cytochrome c Standard with deionized or distilled water. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions.
Protocol tips
-When mixing or reconstituting protein solutions, always avoid foaming. -Substrate Solution should remain colorless until added to the plate. -Keep Substrate solution protected from light. -Stop Solution should be added to the plate in the same order as the Substrate Solution. -Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the Substrate Solution.

Publication protocol

Plasma levels of Angiopoietin‐2 (MANG20, R&D Systems) and Endocan‐1 (MBS039900, MyBioSource, San Diego, CA) were measured as markers of endothelial permeability by ELISA according to the manufacturer's protocol. Plasma, kidney, and lung IL‐1β (RLB00, R&D Systems) and IL‐18 (ab213909, Abcam, Boston, MA) were quantified by ELISA according to the manufacturer's protocol. Levels of IL‐1β in collected platelet media were also quantified by ELISA according to the manufacturer's protocol. Kidney and lung lysates were prepared with the BioPlex Cell Lysis Kit (171304011, BioRad, Hercules, CA) according to the manufacturer's protocol. Tissue cytokine levels were normalized to protein concentration using a protein assay with BSA standards (Pierce, Rockford, IL).

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Manufacturer protocol

Download the product protocol from R&D Systems for Mouse/Rat Angiopoietin-2 Quantikine ELISA Kit below.

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