Mouse/Rat Leptin 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

To investigate changes in leptin protein expression due to aging, ELISA was performed on tissue obtained from rats separated into the same two age groups as the above experiments: the young group (10 weeks) and aged group (48 weeks) (n = 10 each).

Quadricep muscles harvested from rats as described above were homogenized in radioimmune precipitation (RIPA) buffer (Wako Pure Chemical Co., Inc., Osaka, Japan) containing an added protease inhibitor cocktail (Roche, Madison WI, USA). Total protein concentration in the solution was ascertained by the bicinchoninic acid (BCA) assay (Thermo Fisher Scientific, Rockford IL, USA) and leptin protein concentration by a rat leptin ELISA kit (R&D Systems, Inc., Minneapolis MN, USA).

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Manufacturer protocol

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

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