Tail vein blood was collected on the second diestrus day of the estrous cycle [24 (link)] during the week prior to OVX, at 5 weeks post-OVX, and again at the time of sacrifice. Blood was drawn during the latter part of the light cycle; plasma was isolated and stored at −80 °C until analyzed. Concentrations of insulin, leptin, amylin, and glucagon were simultaneously measured using the Rat Endocrine LINCOplex Kit 96 Well Plate Assay (RENDO-85 K; Millipore, St Charles, MO). Colorimetric assays were used to measure plasma free fatty acids (Wako Chemicals USA, Richmond, VA), glucose, triglycerides (TG), and total cholesterol (#TR15421, TR22321, and TR13521, respectively; Thermo Fisher Scientific, Waltham, MA).
Tr15421
Manufactured by Thermo Fisher Scientific
Sourced in United States
The TR15421 is a laboratory centrifuge designed for general-purpose applications. It features a maximum rotation speed of 15,000 rpm and a maximum relative centrifugal force of 21,420 x g. The centrifuge is equipped with a brushless motor and a microprocessor control system for precise speed and time settings.
Automatically generated - may contain errors
Lab products found in correlation
Tr22321, by Thermo Fisher Scientific (5 mentions)
Tr13521, by Thermo Fisher Scientific (4 mentions)
Tr13421, by Thermo Fisher Scientific (2 mentions)
Tr22421, by Thermo Fisher Scientific (2 mentions)
Onetouch ultraeasy, by Johnson & Johnson (2 mentions)
22 lepms e01, by ALPCO (1 mentions)
Mak006, by Merck Group (1 mentions)
Khb3481, by Thermo Fisher Scientific (1 mentions)
Khb3441, by Thermo Fisher Scientific (1 mentions)
Kho0631, by Thermo Fisher Scientific (1 mentions)
Mak043, by Merck Group (1 mentions)
Mouse rat leptin elisa, by ALPCO (1 mentions)
Rat insulin elisa, by ALPCO (1 mentions)
Mk118, by Takara Bio (1 mentions)
80 insrt e01, by ALPCO (1 mentions)
10 protocols using tr15421
1
Endocrine Profiling in Ovariectomized Rats
2
Blood Biomarker Measurement Protocol
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Blood was collected from the inferior vena cava at the time of euthanasia, and serum was stored at −80°C until analysis. All analyses were performed in duplicate. Plasma insulin and leptin were measured by ELISA (80‐INSRT‐E01 and 22‐LEPMS‐E01, respectively, ALPCO, Salem, NH). Limits of detection (LOD) were 0.124 ng/mL for insulin and 10 pg/mL for leptin. Colorimetric assays were used to measure plasma nonesterified fatty acids (NEFA; Wako Chemicals USA, Richmond, VA), glucose, triglycerides (TG), and total cholesterol (#TR15421, TR22321, and TR13521, respectively; Thermo Fisher Scientific, Waltham, MA).
3
Biochemical Assays for Livestock Research
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Concentrations of progesterone for Experiment 1 were determined utilizing a commercially available RIA kit (207270; MP Biomedicals; Irvine, CA, USA) according to manufacturer’s recommendations. Intra-assay coefficients of variation averaged 6.39% and inter-assay coefficients of variation were 7.25% for the analysis of samples containing 0.5, 1.0, and 5.0 ng progesterone/mL. Concentrations of FGF21 (RD291108200R; BioVendor, LLC; Asheville, NC, USA), progesterone (Experiment 2; 4825; Monobind Inc.; Lake Forest, CA, USA), NEFA (999-34691; Wako Diagnostics; Terra Bella, CA, USA), PUN (MAK006; Sigma-Aldrich; St. Louis, MO, USA), and glucose (TR15421; Thermo Fisher Scientific; Middletown, VA, USA) were determined utilizing commercially available ELISA kits according to manufacturer’s recommendations. Intra-assay coefficients of variation averaged 3.64, 5.71, 5.26, 1.42, and 5.21% and inter-assay coefficients of variation were 7.53, 14.69, 6.49, 4.21, and 5.95% for analysis of samples containing 25.6 pmol FGF21/L, 0.64 ng progesterone/mL, 0.67 mmol NEFA/L, 4.8 mmol PUN/L, and 9.3 mmol glucose/L, respectively. Recovery of the added mass of 8 replicates each containing 100 pg of recombinant bovine FGF21 (cyt-657; Prospec; Ness Ziona, Israel) resulted in 115.65% recovery.
4
Plasma Biomarker Panel in Lean Individuals
5
Plasma Biomarkers Evaluation in Rodents
6
Plasma Biomarkers in Ovariectomized Rats
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Tail vein blood was collected at the time of OVX, at 2 weeks post-OVX, and again at the time of sacrifice. Blood was drawn during the latter part of the light cycle; plasma was isolated and stored at − 80 °C. Plasma insulin was measured by ELISA (Alpco 80-INSRT-E01, Salem, NH). Colorimetric assays were used to measure plasma-free fatty acids (Wako Chemicals USA, Richmond, VA), glucose, triglycerides (TG), and total cholesterol (#TR15421, TR22321, and TR13521, respectively; Thermo Fisher Scientific, Waltham, MA). Inflammatory markers were measured using a 90-plex antibody array (Rat L90 Array, AAR-SERV-LG, RayBiotech Life, Inc., Peachtree Corners, GA) and pathway analysis was performed using Enrichr software [43 (link)].
7
Hormone Profiling in Euthanized Rats
8
Quantifying Serum Metabolic Markers
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Colorimetric assays were used to measure serum non-esterified free fatty acids (NEFA) (Wako Chemicals USA, Richmond, VA), glucose and triglycerides (TAGs), (TR15421 and TR22321, respectively, Thermo Fisher Scientific, Waltham, MA). Concentrations of insulin and leptin were simultaneously measured in serum using the Milliplex Map Mouse Serum Adipokine Immunoassay (MADPK-71k-04, Millipore, Billerica MA).
9
Glucose and Insulin Tolerance Tests
10
Metabolic Profiling of HFD Mice
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GTT and ITT were performed as described previously (Xu et al., 2019 (link)). In brief, for GTT, 6‐h fasted mice were injected i.p. with d ‐glucose (1 g·kg−1), and blood samples were collected and measured at 0, 30, 60, 90, and 120 min after glucose injection. For ITT, 4‐h fasted mice were given insulin (1 U·kg−1), and blood samples were collected and measured at the same time points as GTT with a glucometer (OneTouch UltraEasy, Johnson & Johnson). Body composition of mice was assessed by NMR (MiniSpec LF90II TD‐NMR Analyzer). Oxygen consumption (VO2), carbon dioxide production (VCO2), food intake, and heat production were monitored with CLAMS (Columbus Instruments). At the end of HFD administration, mice were anaesthetised with ketamine (80 mg·kg−1) and xylazine (5 mg·kg−1), given i.p.. Mice were fasted for 12–14 h before blood was collected. Individual kits were purchased from Thermo Scientific for measuring serum concentrations of cholesterol, triglyceride, and glucose (#TR15421, TR13421, and TR22421; Thermo Scientific). The content of hepatic triglyceride was determined using tissue lipids extracted by a mixture of chloroform and methanol as previously described (Xu et al., 2019 (link)).
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