Mouse TNF-alpha 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

Cell-free BAL fluid was evaluated for cytokines and chemokines by ELISA. Levels of mediators previously implicated in mediating ODE-induced airway inflammation include TNF-α, IL-6, and the murine neutrophil chemoattractants, CXCL1 and CXCL2, [15] were quantitated by Quantikine ELISAs from R&D Systems (Minneapolis, MN) according to manufacturer’s instructions. Levels of cytokines classically associated with allergic disease including IL-4, IL-5, and IL-13 were quantitated using the Ready-SET-go ELISA kits (Affymetrix, Thermo Fisher). Levels of chemokines including CCL2, CCL3, CCL8, CCL11, CCL12, and CCL22, and levels of cytokines including IL-9, IL-17A, IL-17E, and IL-33, were quantitated by murine Luminex multiplex assay from R&D Systems according to manufacturer’s instructions.

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Manufacturer protocol

Download the product protocol from R&D Systems for Mouse TNF-alpha Quantikine ELISA Kit below.

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