Rat ICAM-1/CD54 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

Quantitative assay for ET‐1 and cytokines (TNFα, IL‐6, sICAM‐1 and sVCAM‐1) in the supernatants or in rat serum was performed using the Quantikine ELISA Kits according to the manufacturer's instruction. Briefly, 50 μL serum or cell supernatant was added to a 96‐well polystyrene microplate pre‐coated with various monoclonal antibodies against the corresponding cytokines and incubated for 2 hours at room temperature on the shaker. After five times of gentle washes with PBS, 100 μL secondary antibodies conjugated to horseradish peroxidase were added and incubated for 2 hours at room temperature, followed by an addition of 100 μL substrate solutions and incubation for 30 minutes at room temperature. Then, 100 μL stop solution was added and the optical density (OD) was read at 450 nm in a microplate reader (Bio‐Rad Laboratories, Hercules, USA).

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Manufacturer protocol

Download the product protocol from R&D Systems for Rat ICAM-1/CD54 Quantikine ELISA Kit below.

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