Terminal bleeds from mice were incubated at room temperature for 1 hour in serum separator tubes (Sarstedt, Nümbrecht, Germany) and then spun at 10,000 × rpm for 5 minutes. Serum was collected and stored at −80°C until analysis. Mouse leptin, human NAG-1, mouse IGF-1 (R&D, Minneapolis, MN), mouse insulin (Alpco, Salem, NH), and mouse glycerol (Cayman, Detroit, MI) ELISA kits was used according to manufacturer’s instructions. Cholesterol and triglycerides kit was purchased from Beckman Coulter (Melvill, NY) and non-esterified free fatty acid kit was purchased from Sekisui Diagnostics (Exton, PA). Analysis was done using an Olympus AU400e clinical analyzer by Beckman Coulter.
Au400e clinical analyzer
Manufactured by Beckman Coulter
The AU400e is a clinical analyzer designed for automated biochemical analysis. It performs quantitative tests on a variety of biological samples, including serum, plasma, and urine. The AU400e is capable of processing a wide range of diagnostic tests, providing accurate and reliable results to support patient care.
Automatically generated - may contain errors
Lab products found in correlation
Non esterified free fatty acid kit, by Sekisui (4 mentions)
Serum separator tubes, by Sarstedt (2 mentions)
Mouse glycerol, by Cayman Chemical (2 mentions)
Human nag 1, by R&D Systems (2 mentions)
Mouse igf 1, by R&D Systems (2 mentions)
Cholesterol and triglycerides kit, by Beckman Coulter (2 mentions)
Humulin r, by Eli Lilly (1 mentions)
5 protocols using au400e clinical analyzer
1
Serum Biomarker Measurement in Mice
2
Glucose and Insulin Metabolism Assessment
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Plasma insulin levels were determined by ELISA (Crystal Chemistry for ob/ob mice; ALPCO Diagnostics for high-fat diet–fed mice). The glucose × insulin product was calculated by multiplying 5-h food-removed glucose and insulin values. Cholesterol measurements in high-fat diet–fed mice were determined on an AU400E Clinical Analyzer (catalog #OSR6283; Beckman Coulter). For glucose and insulin tolerance tests, ob/ob mice were fasted for 5 h and injected with glucose (1 g/kg, s.c.) or insulin (2.4 units/kg, s.c.; Humulin R; Eli Lilly), and high-fat diet–fed mice were fasted for 3.5–4 h and injected with insulin intraperitoneally.
3
Serum Biomarker Measurement in Mice
4
Bone and Brain Analyses in Rats
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Following completion of MWM testing, samples were collected from rats randomly selected from shipments 3 and 4. Blood was collected via a cardiac puncture from rats deeply anesthetized under CO2 and plasma separated by centrifugation in EDTA-free, heparin coated plasma tubes (# 367874, Becton Dickinson, Franklin Lakes, NJ). The brain and femur were excised, immediately frozen on dry ice, and stored at -80 °C. Cleaned femur samples (5–8 mg) were ashed (8 h; 590 °C), pulverized, and weighed. Brain samples were homogenized in 3-ml di-H2O. For PND25 assessment, the brain and femur samples were obtained from randomly selected unassigned male rats. Duplicate samples were analyzed using a modification of the hexamethyldisiloxane microdiffusion method of Taves and Neuman (1964 (link)) and detected with a fluoride ion-specific electrode and a pH/ISE meter. Urine was collected in metabolism cages (Techniplast) from individual rats (n = 10) in G2 and G4 between 9:00–13:00 h and frozen. No water was provided over this interval. Urine samples < 200 μl were excluded. A 100-μl aliquot of urine was analyzed for creatinine using a kinetic modification of the Jaffe procedure (Moore and Sharer 2017 (link)). The rate of change at 520/800 nm was determined using the Olympus AU400e clinical analyzer (Beckman-Coulter Irving, TX).
5
Mouse Tamoxyfen Diet and Blood Monitoring
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Animal experiments were conducted according to American Association for the Accreditation of Laboratory Animal Care guidelines and were approved by the institutions Animal Welfare Committee (Cold Spring Harbor Laboratory's Institutional Animal Care and Use Committee guidelines). Mice were maintained at a constant temperature of 23°C and were allowed standard lab diet and water ad libitum. As indicated, male mice were given a tamoxifen diet (Harlan). Body weights were recorded weekly. Blood was collected weekly or bi-weekly by tail bleed, and plasma was separated by centrifugation at 10 000 x g for 10 min at 4°C. Blood chemistries were determined on an AU400E Clinical Analyzer (Beckman Coulter).
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