Human CCL2/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

The MCP-1 levels in the plasma samples were analyzed using human MCP-1 enzyme-linked immunosorbent assay (ELISA) kits (R&D Systems, Abingdon, UK) as previously described [56]. Briefly, the wells of an ELISA plate containing 0.1 mL of diluted samples were incubated at 37 °C for 2 h and, subsequently, 0.1 mL of anti-human MCP-1 antibody was added to each well. Each well was washed three times with 0.4 mL washing buffer, and then 0.2 mL of conjugate solution was added. Again, the wells were washed and loaded with 0.2 mL of TMB agent at 37 °C in a dark room for 30 min. For final processing, each well was filled with 0.1 mL of stop solution, and the amount of MCP-1 absorbance was quantitatively measured at 450 nm with a spectrophotometer and calibrated using human MCP-1 standards.

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Manufacturer protocol

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

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