Rat Leptin ELISA Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Bring all reagents and samples to room temperature (18–25 °C) before use.	-Wash by filling each well with Wash Buffer (300 µl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels. -Read at 450 nm immediately after adding STOP solution.

Upstream tips
-Bring all reagents and samples to room temperature (18–25 °C) before use.
Protocol tips
-Wash by filling each well with Wash Buffer (300 µl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels. -Read at 450 nm immediately after adding STOP solution.

Publication protocol

Blood glucose was measured using OneTouch Ultra2 blood glucose meter (Life Scan, Milpitas, CA). Serum β-hydroxybutyrate was determined using the Cayman Chemical β-hydroxybutyrate assay kit (Ann Arbor, MI). Serum triglyceride (TG) and cholesterol were measured using the Infinity Assay Reagents (Thermo Scientific, Waltham, MA). Serum free fatty acids (FFA) were determined with a BioVision FFA Kit (Milpitas, CA). Activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured with Infinity kit (Thermo Scientific). Serum adiponectin and leptin levels were determined using ELISA kits (Millipore, Billerica, MA). Serum ethanol concentration was measured using BioVision ethanol assay kit.

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Rat Leptin ELISA Kit below.

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