Mouse CCL2/JE/MCP-1 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

In this study, urine and kidney lysate CCL2, CCL5, and CXCL9 levels were measured by using ELISA kits from R&D Systems (Minneapolis, MN, USA) in accordance with the instructions of the manufacturer. Briefly, diluted urine or kidney lysate samples were added to capture antibody pre-coated 96-well microplates. After incubation with samples, the detection antibody was added, followed by streptavidin-HRP, and substrate. A microplate reader ELX808 from BioTek Instruments (Winooski, VT, USA) was used to read the optical density at 450 nm. The concentration was calculated based on a standard curve. Urinary creatinine concentrations were determined by using Creatinine Parameter Assay Kit (R&D Systems). Urine creatinine concentrations were used to normalize urine chemokine concentrations.

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Manufacturer protocol

Download the product protocol from R&D Systems for Mouse CCL2/JE/MCP-1 Quantikine ELISA Kit below.

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