IPGTT's were carried out according to the IMPReSS protocols (https://www.mousephenotype.org/impress/protocol/87/8 ) using 2 g glucose per kg body weight. Mice were fasted overnight and blood sampled under a local anesthetic at 0 min (baseline) and 60 and 120 min post glucose injection. Whole blood glucose was measured using an AlphaTRAK meter and test strips (Abbott Animal Health, UK). At the 13 week time point male plasma samples were also run on a GM9 glucose analyzer (Analox Instruments, UK) which is more sensitive than the AlphaTRAK meter. Plasma insulin was assayed using a Mouse insulin ELISA kit (Mercodia, Sweden).
Gm9 glucose analyzer
Manufactured by Analox Instruments
Sourced in United States, United Kingdom
The GM9 is a glucose analyzer designed for laboratory use. It provides accurate measurements of glucose levels in samples.
Automatically generated - may contain errors
Lab products found in correlation
Mouse insulin elisa kit, by Mercodia (1 mentions)
Alphatrak meter and test strips, by Abbott (1 mentions)
Radioimmunoassay kit, by Merck Group (1 mentions)
Cortisol, by MP Biomedicals (1 mentions)
14c oa, by PerkinElmer (1 mentions)
Beckman scintillation counter, by PerkinElmer (1 mentions)
Colorimetric assay kit, by Fujifilm (1 mentions)
Mouse ultrasensitive insulin elisa kit, by ALPCO (1 mentions)
Cobas c111 analyzer, by Roche (1 mentions)
6460 triple quadrupole, by Agilent Technologies (1 mentions)
Fp 6200, by Jasco (1 mentions)
Enzyme linked immunosorbent assay, by R&D Systems (1 mentions)
Pi 12k, by Merck Group (1 mentions)
Radioimmunoassay, by Merck Group (1 mentions)
Onetouch ultra2, by LifeScan (1 mentions)
Monarch chemistry analyzer, by Werfen (1 mentions)
Dade behring dimension rxl chemistry analyzer, by Siemens (1 mentions)
13 protocols using gm9 glucose analyzer
1
Glucose Tolerance in Mice
2
Comprehensive metabolic profiling in research
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Blood samples were collected and processed as previously described (68 (link)). Plasma glucose was analyzed using the glucose oxidase method (GM9 Glucose Analyzer, Analox Instruments). Insulin and glucagon were assayed using commercially available radioimmunoassay kits (Millipore). Cortisol (MP Biomedicals) and GH (Thermo Fisher Scientific) were assayed using commercially available ELISA kits. NEFAs were measured using a fluorometric assay (Sigma-Aldrich). Catecholamines and amino acids were assessed using high-performance liquid chromatography and glucose tracer enrichment was assessed using gas chromatography/mass spectrometry.
3
Quantifying Lactobacillus Fatty Acid Uptake
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Fatty acid consumption of Lactobacillus strains, L. rhamnosus GG (ATCC 53103), L. acidophilus (ATCC 4356), and L. gasseri (ATCC 33323) in bacterial growth medium was evaluated quantitatively using radioactive tracers, [14C]-OA and [14C]-palmitic acid (PA) (PerkinElmer, Waltham, MA). The glucose concentration in each bacterial medium was measured using a GM9 glucose analyzer (Analox Instruments, London, UK). Overnight cultures of Lactobacillus were diluted 100-fold (v/v) and subcultured three times to achieve viability, as described previously27 (link). [14C]-OA (1 µci) was added to 10 ml of MRS growth medium at 20 g/l, and three Lactobacillus strains (1x108 cfu of each strain) were cultured for 6 h with shaking (37 °C at 220 rpm). For L. rhamnosus GG, 1 μci of [14C]-OA and [14C]-PA were further tested. One milliliter of cultured broth was aliquoted every 3 h and centrifuged (10 min, 4000×g), and the supernatant was collected. The pellet was resuspended and washed three times in MRS broth (10 min, 4000×g). 14C radioactivity was measured in both the supernatant and pellet of each aliquoted sample using a β-counter (Beckman scintillation counter, PerkinElmer, Waltham, MA).
4
Glucose Tolerance Test in Fasted Rats
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Rats were fasted (food removed at 9:00 AM) for 24 h prior to IVGTT, and IVGTT was performed as described [9 (link)] (Supplementary Figure 1a ). Blood samples were collected in heparinized tubes and centrifuged at 2,000×g for 30 s. Plasma glucose levels were determined by the glucose oxidase method using a GM9 glucose analyzer (Analox Instruments, Stourbridge, UK).
5
Metabolic Biomarker Quantification Protocol
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Blood glucose during hyperinsulinemic-clamp and glucose tolerance test was measured using the GM9 Glucose Analyzer (Analox Instruments). Lipids in tissue were extracted using the method of Bligh and Dyer55 (link) and hepatic triglyceride, cholesterol and plasma NEFA were measured by colorimetric assay kits from Wako (Wako Pure Chemical Industries, 279-75401). Plasma triglyceride, total cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL), ALT, and AST were measured using a Cobas c111 analyzer (Roche). Plasma insulin was measured using Mouse Ultrasensitive Insulin ELISA kits (ALPCO, 80-INSMSU-E01).
6
Plasma Glucose and BCAA Kinetics Analysis
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Plasma glucose was measured by the glucose oxidase method (GM9 glucose analyzer, Analox Instruments, Lunenburg, Massachusetts, USA). Total BCAA concentrations were measured during clamp by a 4-min enzymatic, fluorimetric method (FP-6200; Jasco Corporation, Tokyo, Japan).24 (link) Glucose turnover was calculated as in Ref. 24 (link). Serum insulin was measured by ELISA (Mercodia AB, Uppsala, Sweden), and non-esterified fatty acid (NEFA) concentrations by colorimetric assay (NEFA-HR, Wako Chemicals USA, Richmond, VA, USA). Expired air was analyzed for 13CO2 enrichment by isotope ratio mass spectrometry (Micromass 903D, Vacuum Generators, Winsforce, UK). Individual plasma AA and the stable isotopic enrichments of the tracers were measured by liquid chromatography-tandem mass spectrometry (Agilent 6460 Triple Quadrupole, with UHPLC 1290, Santa Clara, CA, USA). l -[1–13C]Leucine kinetics were calculated at the isotopic steady states, according to Ref. 29 (link), using plasma [1–13C]α-ketoisocaproic acid (KIC) enrichment (reciprocal model).
7
Biomarker Sampling and Analysis
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Samples used to measure glucose were processed using GM9 glucose analyzer (Analox Instruments, Atlanta, GA). Samples for insulin, adiponectin, and leptin were drawn into ethylenediamine tetraacetic acid (EDTA) tubes, centrifuged within 10 to 15 minutes at 3000 RPM, and stored at −80°. Insulin and leptin samples were analyzed at Vanderbilt’s Hormonal Lab Core. High-molecular-weight adiponectin and interleukin 6 (IL-6) were measured by enzyme-linked immunosorbent assay (R&D Systems, Minneapolis, MN) and high-sensitivity C-reactive protein (hsCRP) was measured at the Vanderbilt Clinical laboratory by immunoturbidimetric method.
8
Serum and Urine Osmolality Measurement
9
Liver Metabolic Profiling in Humans
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Arterial, hepatic portal vein, and hepatic vein whole blood samples collected during the study were analyzed using a variety of standard methods to obtain data regarding hormone and substrate balance across the liver. The arterial and portal blood samples were collected at the same time, followed by the hepatic venous blood sample. This allowed for the transit time of blood across the liver. Plasma glucose samples were immediately measured upon collection using a GM9 glucose analyzer (Analox Instruments Ltd., Stourbridge, UK). Blood lactate, glycerol, alanine, and non-esterified fatty acids were analyzed using enzymatic spectrophotometric methods (see supplement ). Plasma insulin (#PI-12K, MilliporeSigma, Burlington, MA), glucagon (#GL-32K, MilliporeSigma), and cortisol (VUMC Analytical Services in-house primary antibody with I125 cortisol from MP Biomedicals, Santa Ana, CA) were measured by radioimmunoassay. All samples were kept in an ice bath during the experiment and subsequently stored at −80°C until assays were performed. For the determination of [3-3H]-glucose, plasma samples were deproteinized and quantified using liquid scintillation counting (40 (link)). Hepatic glycogen was quantitatively assessed using the amyloglucosidase method (see supplement ).
10
Plasma Glucose and Insulin Measurement
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Plasma glucose concentrations were measured by the glucose oxidase method (Glucose Analyzer GM9, Analox Instruments, Lunenburg, MA, USA). Plasma insulin levels were determined by radioimmunoassay (Millipore Canada Ltd, Etobicoke, ON, Canada).
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