Blood samples were drawn in the morning, after an over-night fasting. Venous plasma (p)-glucose (mmol/L) was analyzed with a blood glucose analyzer (Hemocue AB, Ängelholm). P-Insulin (mIE/L) was measured with a competitive radioimmunoassay with intra and interassay CVs of 7,1% or less. Serum (s)—IGF-1 was measured with a chemiluminescent immunoassay. The normal range was 71–239 μg/L in subjects aged 31–42 years (interassay coefficient of variation (CV)%, 8% at the level of 30 μg/L and 8% at the level of 239 μg/L). The homeostasis model of assessment (Homa) index was calculated as the product of the fasting p-insulin level (microU/mL) and the fasting p-glucose level (mmol/L), divided by 22.5. Leptin was measured with a commercially available RIA-kit from Millipore Corporation, Linco Research, Inc. The within, between and total CV% were determined to be approximately 4.5%, 2.3% and 5.1% respectively at 3.9 μg/L. The corresponding figures at 20.5 μg/L were 5.1%, 2.1% and 5.5%. Ghrelin (total) was measured with a commercially available RIA-kit from Millipore Corporation, Linco Research, Inc. The analytical sensitivity of the method was estimated to be typically better than 100 ng/L. The within, between and total CV% were determined to be approximately 6.4%, 2.5% and 6.9% respectively at 550 ng/L. The corresponding figures at 1229 ng/L were 3.9%, 5.7% and 6.9%.
Ria kit
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom
The RIA kit is a laboratory equipment product designed for radioimmunoassay (RIA) analysis. RIA is a sensitive analytical technique used to measure the concentration of specific substances, such as hormones, drugs, or other analytes, in a sample. The RIA kit provides the necessary components and materials to perform this type of analysis in a laboratory setting.
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Lab products found in correlation
Glucagon, by Merck Group (2 mentions)
Mouse crp elisa kit, by Thermo Fisher Scientific (2 mentions)
Onetouch ultra2, by LifeScan (2 mentions)
Dextrose, by Merck Group (2 mentions)
Dpc immulite analyzer, by Diamond Diagnostics (2 mentions)
Accu check go, by Roche (1 mentions)
Accucheck 2, by Roche (1 mentions)
Ultra sensitive mouse insulin elisa kit, by ALPCO (1 mentions)
Tina quant hba1c gen 3 kit, by Roche (1 mentions)
Elisa immunoassay kit, by Merck Group (1 mentions)
Elisa assay kit, by Merck Group (1 mentions)
Trasylol, by Bayer (1 mentions)
Hlc 723g7 hemoglobin hplc analyzer, by Tosoh (1 mentions)
Cobas 8000, by Roche (1 mentions)
Cobas e801, by Roche (1 mentions)
Dimension vista 1500 system, by Siemens (1 mentions)
Cobas c111 system, by Roche (1 mentions)
Glucometer, by Arkray (1 mentions)
C peptide, by ALPCO (1 mentions)
Gdf 15, by Merck Group (1 mentions)
51 protocols using ria kit
1
Metabolic Biomarker Assessment Protocol
2
Insulin Measurement by RIA Kit
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Insulin was measured by a RIA kit (Millipore, Missouri, U.S.A.), according to manufacturer’s protocol.
3
Glucose Tolerance and Insulin Levels
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Food was removed from cage hoppers at 08:30 h, and 6 h later a dose of glucose (1 g/kg body weight) was injected into the peritoneal cavity. Tail vein blood was collected at 0, 15, 30, 45, 60 and 90 min after the glucose bolus injection for the determination of glucose concentrations using a glucometer (Accu-Check Go, Roche) and insulin levels using a RIA kit from Millipore (Millipore, Billerica, MA, USA). Glucose tolerance curves for glucose and insulin are presented as absolute values. Area under the glucose or insulin concentration curves between 0 and 90 min after glucose ingestion were calculated, subtracting glucose or insulin concentrations prior to glucose ingestion.
4
Insulin Secretion and Sensitivity Assessment
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Serum insulin, C-peptide and leptin levels were measured by the RIA kit, according to the manufacturer’s protocol (Millipore, Billerica, MA, USA). Insulin:C-peptide ratio (insulin and C-peptide are secreted in equimolar quantities by the β-cells; an insulin:C-peptide ratio >1 may therefore indicate reduced insulin clearance), glucose-to-insulin ratios, homeostatic model assessment-insulin resistance (HOMA-IR) index and the quantitative insulin sensitivity check index (QUICKI) were calculated as previously defined [63 (link)].
5
Cellular Insulin Quantification Protocol
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Cells were seeded in six-well culture dishes and grown in complete medium. The medium and cell lysates were harvested at the indicated time points. The incubation medium was gently centrifuged to remove floating cells. The cell density was determined after trypsinization in a Neubauer counting chamber. For the measurement of cellular insulin content, the cells were homogenized in acetic acid. Insulin content and secretion levels were measured using a commercially available RIA kit (Millipore) according to the manufacturer’s protocol.
6
Insulin Secretion Assay in beta Cells
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βTC-tet cells or Min6(B1) were transfected with siCT or siClic4. 72 h later, they were washed with PBS and pre-incubated for 2 h in KRBH-BSA (Krebs-Ringer bicarbonate HEPES buffer containing 120 mM NaCl, 4 mM KH2PO4, 20 mM HEPES, 1 mM MgCl2, 1 mM CaCl2, 5 mM NaHCO3, and 0.5% BSA, pH7.4) supplemented with 2 mM glucose, later the medium was replaced with fresh KRBH-BSA containing 2 or 20 mM glucose and incubated for 1 h in the presence or absence of 100 nM Exendin-4. Secreted and cellular insulin were assessed by radioimmunoassay (RIA) using RIA kit (Millipore, MA, USA) following manufacturer's instructions.
7
Comprehensive Metabolic Profile Analysis
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The heparinized blood was centrifuged at 3,500×g at 4°C for 15 min. Glucose levels, total lipid profile, uric acid, urea, bilirubin, electrolytes, iron, and the activities of alanine aminotransferase (ATL), aspartate aminotransferase (AST), and alkaline phosphatase (AP) were determined in the harvested plasma by utilizing automated systems in the Department of Clinical Chemistry of the UMG. In addition, serum leptin levels were measured with a radio-immuno-assay (RIA) kit (Millipore, MA, USA) for all groups. Serum C-peptide and insulin concentrations were evaluated in the Co, LDC, and L-EF groups with RIA (Millipore, MA, USA). The overnight-fasted animals were tested for plasma glucose and TG, and serum insulin and C-peptide levels.
8
Insulin Secretion Regulation by Glucose
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The cultured cells in all groups were exposed to glucose-
free Krebs-Ringer bicarbonate (KRB, Sigma-Aldrich,
USA) for 1 hour. Then, the cells of each group were
divided in three groups and exposed to KRB containing
glucose at the concentration of 5.56, 16.7 and 25 mmol/L
for 1 hour, Insulin contents were determined using a RIA
kit (Millipore, Germany) (5 (link), 8 (link)).
free Krebs-Ringer bicarbonate (KRB, Sigma-Aldrich,
USA) for 1 hour. Then, the cells of each group were
divided in three groups and exposed to KRB containing
glucose at the concentration of 5.56, 16.7 and 25 mmol/L
for 1 hour, Insulin contents were determined using a RIA
kit (Millipore, Germany) (5 (link), 8 (link)).
9
Insulin and Glucose Tolerance Testing
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Insulin tolerance tests were performed on male mice at 15 weeks of age (unless otherwise stated) at 14:00–16:00 h. Briefly, the mice were fasted for 6 h and then intraperitoneally injected with insulin (0.5 IU/kg) (Novo Nordisk Pharmaceuticals, Baulkham Hills, NSW, Australia). Tail tip blood samples were then collected at 0, 15, 30, 45, 60, 75 and 90 min after insulin injection, and blood glucose levels were measured using a glucometer (AccuCheck II; Roche, New South Wales, Castle Hill, Australia). 7 days later (unless otherwise stated), mice underwent a glucose tolerance test, where they were fasted for 16–24 h before intraperitoneal injection between 13:00 and 15:00 h of a 10% d -glucose solution (1.0 g/kg) (Astra Zeneca, North Ryde, NSW, Australia). Blood samples were obtained from the tail tip at 0, 20 and 60 min after glucose injection and blood glucose levels were measured using a glucometer. Serum was also separated, immediately frozen and stored at −20 °C for later measurement of serum insulin using an ELISA kit from Linco Research (St Charles, Missouri, USA) or a RIA kit from Millipore (Millipore, Billerica, MA, USA).
10
Glucose-stimulated C-peptide and Insulin Secretion
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Cultured cells were washed in Krebs-Ringer-bicarbonate (KRB) washing buffer (130 mM NaCl, 3.6 mM KCl, 1.5 mM CaCl2, 0.5 mM MgSO4, 0.5 mM KH2PO4, 2.0 mM NaHCO3, and 10 mM HEPES) and incubated in KRB buffer containing 1 mM glucose for 1 h. The cells were then stimulated for 1 h in KRB buffer containing 25 mM glucose. C-peptide and insulin concentrations were measured with a RIA kit (Millipore).
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