Glucose liquicolor assay kit

Field formulation TFM (Clariant SFC GMBH WERK, Griesheim, Germany) was used for all experiments [35% active ingredient dissolved in isopropanol; provided courtesy of Fisheries and Oceans Canada (DFO), Sea Lamprey Control Centre, Sault Ste. Marie, ON, Canada], and TFM exposure concentrations were verified using precision TFM standards provided by DFO. The Glucose Liquicolor® assay kit (Stanbio Laboratory, Boerne, TX, USA) was used to measure tissue and plasma glucose levels. High-binding 96 well plates (Immulon HB, VWR Canlab, Mississauga) were used for an in-house cortisol enzyme-linked immunosorbent assay (ELISA), used in conjunction with a cortisol monoclonal antibody conjugated to horseradish peroxidase (East Coast Bio, ME, USA). The RiboZol^TM used for RNA extraction was obtained from AMRESCO (Solon, OH, USA), while the DNaseI was purchased from Sigma-Aldrich and the first strand cDNA synthesis kit was from Invitrogen (Burlington, ON, CA). The PerfeCTa® SYBR® Green FastMix® green fluorescent dye used for real-time quantitative PCR (RT qPCR) was purchased from Quanta Biosciences (Gaithersburg, MD, USA), while the 96-well plates were obtained from Bio-Rad (Mississauga, ON, CA). All other chemicals were purchased from Sigma-Aldrich (MO, USA).

Tissue and plasma glucose levels were measured colorimetrically using the Glucose Liquicolor assay kit (Kit No. 1070–125, Stanbio Laboratory, Boerne, TX, USA), by following the manufacturer’s instructions. Liver protein was determined using the bicinchoninic acid (BCA) method [31 (link)] on the 96-well plate spectrophotometer, with bovine serum albumin as the standard, using a commercial kit (G-Biosciences, Geno Technology, Inc., St. Louis, MO, USA). Plasma lactate levels were determined enzymatically (lactate dehydrogenase), as previously described [12 (link)].