Radioimmunoassay

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom, United States, Belgium

Radioimmunoassay is an analytical technique used to measure the concentration of specific substances, such as hormones, drugs, and other biomolecules, in a sample. It relies on the principle of competitive binding between a radioactively labeled analyte and the unlabeled analyte in the sample, to a limited number of high-affinity antibody binding sites. The amount of radioactive analyte bound to the antibody is inversely proportional to the concentration of the unlabeled analyte in the sample.

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Lab products found in correlation

Dimension autoanalyzer, by Siemens (3 mentions) Au2700 analyzer, by Beckman Coulter (2 mentions) Au2700 analyser, by Beckman Coulter (2 mentions) Vacutainer, by BD (1 mentions) Human igf1 elisa kit, by Abcam (1 mentions) Relefact, by Sanofi (1 mentions)

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Radioimmunoassay kits (4 citations) BRAHMS anti-TPOn radioimmunoassay (RIA) kit (2 citations)

9 protocols using radioimmunoassay

Serum Hormone Assay Protocol

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For most participants of the study, venous blood was obtained in the morning and serum was separated immediately.
For the cohort, serum levels of total testosterone were determined by radioimmunoassay (Biosource, Belgium, normal range < 0.17–2.65 nmol/l), estradiol was determined by radioimmunoassay (Immunotech, Czech Republic, normal range 128–541 pmol/l), and SHBG was determined by radioimmunoassay (Immunotech, Czech Republic, normal range 20–85 nmol/l). The intra- and inter-assay coefficients of variation (CVs) for measurement of total testosterone were respectively less than 8% and less than 5%, CVs for estradiol were 7.5% and 13%, and CVs for SHBG were less than 6.7% and less than 8.2%.
Estradiol was measured without regard to the particular phase of menstrual cycle.

Šimonienė D., Platūkiene A., Prakapienė E., Radzevičienė L, & Veličkiene D. (2019). Insulin Resistance in Type 1 Diabetes Mellitus and Its Association with Patient’s Micro- and Macrovascular Complications, Sex Hormones, and Other Clinical Data. Diabetes Therapy, 11(1), 161-174.

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Comprehensive Metabolic Profiling of Participants

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Before surgery and after an overnight fast, blood samples were obtained from the antecubital vein and placed in vacutainer tubes (BD vacutainer™). The serum was separated by centrifugation for 15 min at 4000 rpm and immediately frozen at − 80 °C until analysis. Serum glucose, cholesterol, triglycerides, HDL cholesterol (HDL-C), and C-reactive protein (CRP) were measured in a Dimension autoanalyzer (Dade Behring Inc.) by enzymatic methods (Randox Laboratories Ltd.). LDL cholesterol (LDL-C) was calculated using the Friedewald equation. Insulin was quantified by radioimmunoassay supplied by BioSource International Inc., Camarillo, CA, USA. The homeostasis model assessment of insulin resistance (HOMA-IR) was calculated with the following equation: HOMA-IR = fasting insulin (μIU/mL) × fasting glucose (mmol/L)/22.5. Serum 25(OH)D and parathyroid hormone levels were determined by enzyme immunoassay (ELISA) kits (Immundiagnostik and DRG Diagnostics, respectively). Corrected calcium was calculated using the following equation: fasting calcium (mg/dl) + 0.8 × (4-fasting albumin (g/dl)).

Castellano-Castillo D., Morcillo S., Clemente-Postigo M., Crujeiras A.B., Fernandez-García J.C., Torres E., Tinahones F.J, & Macias-Gonzalez M. (2018). Adipose tissue inflammation and VDR expression and methylation in colorectal cancer. Clinical Epigenetics, 10, 60.

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Comprehensive Metabolic and Hormonal Profile

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Fasting blood samples were obtained from the antecubital vein and placed in vacutainer tubes (BD Vacutainer™, Franklin Lakes, NJ, USA). Serum glucose, total cholesterol, triglycerides, and HDL cholesterol (HDL-c) were measured in a Dimension auto analyzer (Dade Behring Inc., Deerfield, IL, USA) through enzymatic methods (Randox Laboratories Ltd., Crumlin, UK). The LDL cholesterol (LDL-c) was calculated using the Friedewald equation [25 (link)]. Insulin was quantified by radioimmunoassay (BioSource International, Camarillo, CA, USA). Corrected calcium was assessed using a complex metric method from Boehringer Hitachi 717. Alkaline phosphatase was calculated using ELISA (LifeSpan Biosciences Inc., Madrid, Spain). Insulin growth factor type 1 (IGF-1) was determined using Human IGF1 ELISA Kit (Abcam, Madrid, Spain). The homeostasis model assessment of insulin resistance (HOMA-IR) was calculated using the following equation: HOMA-IR = fasting insulin (IU/mL) x fasting glucose (mmol/L)/22.5 [26 (link)].

Cabrera-Mulero A., Crujeiras A.B., Izquierdo A.G., Torres E., Ayers D., Casanueva F.F., Tinahones F.J., Morcillo S, & Macias-Gonzalez M. (2019). Novel SFRP2 DNA Methylation Profile Following Neoadjuvant Therapy in Colorectal Cancer Patients with Different Grades of BMI. Journal of Clinical Medicine, 8(7), 1041.

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Metabolic Biomarkers Assessment

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Serum glucose, cholesterol, triglycerides (TG) and HDL cholesterol (HDL-c) were measured in a Dimension auto analyzer (Dade Behring Inc., Deerfield, IL, USA) by enzymatic methods (Randox Laboratories Ltd., Crumlin, UK). The LDL cholesterol (LDL-c) was calculated from the Friedewald equation. Insulin was quantified by radioimmunoassay (BioSource International, Camarillo, CA, USA). Serum leptin and adiponectin levels were analyzed by enzyme immunoassay (ELISA) kits (respectively: DSL, Webster, FL, USA; and DRG Diagnostics GmbH, Marburg, Germany). The homeostasis model assessment of insulin resistance (HOMA-IR) was calculated from fasting insulin and glucose with the following equation: HOMA-IR = fasting insulin (μIU/mL) × fasting glucose (mmol/L)/22.5 [33 (link)].

Castellano-Castillo D., Moreno-Indias I., Sanchez-Alcoholado L., Ramos-Molina B., Alcaide-Torres J., Morcillo S., Ocaña-Wilhelmi L., Tinahones F., Queipo-Ortuño M.I, & Cardona F. (2019). Altered Adipose Tissue DNA Methylation Status in Metabolic Syndrome: Relationships Between Global DNA Methylation and Specific Methylation at Adipogenic, Lipid Metabolism and Inflammatory Candidate Genes and Metabolic Variables. Journal of Clinical Medicine, 8(1), 87.

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Evaluating Precocious Puberty in Children

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For measuring height, a Harpenden stadiometer (Holdtain Ltd., Crymych, Wales) was used with an increment of 0.1 cm, while weight was measured within 0.1 kg using a CAS scale (CAS, Seoul, Korea). A trained technician performed the measures based on the manufacturer’s guidelines including positioning. BMI calculations and all anthropometry data were used to calculate a standard deviation score (z-score) based on Korean reference data [7 ]. Tanner stage ratings I through V were assigned by a trained rater. Baseline LH, folliclestimulating hormone (FSH), and estradiol (E₂) were collected initially, and then LH and FSH were measured at 30, 45, 60, and 90 minutes after intravenous administration of 100.0 μg gonadorelin acetate (Relefact; Sanofi-Aventis, Frankfurt, Germany). CPP was diagnosed when the peak LH levels after a gonadorelin stimulation was >5.0 U/L. The basal serum E₂ concentration was assayed by a radioimmunoassay (BioSource, Nivelles, Belgium); LH and FSH levels were determined with immunoradiometric assays (BioSource). BA assessment was performed by radiography of the left hand using the Greulich and Pyle method [8 ].

Lee H.Y., Lee Y.J., Ahn M.B., Cho W.K, & Suh B.K. (2018). The effect of overweight on the luteinizing hormone level after gonadorelin stimulation test in girls with idiopathic central precocious puberty. Annals of Pediatric Endocrinology & Metabolism, 23(4), 215-219.

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Comprehensive Metabolic Panel Analysis

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Blood samples were collected and analyzed using the Olympus AU2700 analyzer (Beckman Coulter, High Wycombe, UK) with standard proprietary reagents as follows: glucose with hexokinase, total cholesterol and high-density lipoprotein with cholesterol esterase/oxidase and triacylglycerol with glycerol kinase. Low-density lipoprotein was calculated according to the Friedwald formula. Insulin was measured using radio-immunoassay (Invitrogen, UK). HOMA-IR was calculated using fasting glucose and insulin concentrations.

BOWDEN DAVIES K.A., SPRUNG V.S., NORMAN J.A., THOMPSON A., MITCHELL K.L., HARROLD J.A., FINLAYSON G., GIBBONS C., WILDING J.P., KEMP G.J., HAMER M, & CUTHBERTSON D.J. (2019). Physical Activity and Sedentary Time: Association with Metabolic Health and Liver Fat. Medicine and Science in Sports and Exercise, 51(6), 1169-1177.

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Biochemical Analysis of Blood Samples

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Blood samples were collected and analysed using the Olympus AU2700
analyser (Beckman Coulter, High Wycombe, UK) with standard proprietary
reagents as follows: glucose with hexokinase, total cholesterol and HDL with
cholesterol esterase/oxidase and triacylglycerol with glycerol kinase. LDL
was calculated according to the Friedwald formula. Insulin was measured
using radio-immunoassay (Invitrogen, UK). HOMA-IR was calculated using
fasting glucose and insulin concentrations.

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Metabolic Biomarker Analysis Protocol

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Blood samples were collected and analyzed using the Olympus AU2700 analyzer (Beckman Coulter, High Wycombe, United Kingdom) with standard proprietary reagents as follows: glucose with hexokinase, total cholesterol and HDL-cholesterol with cholesterol esterase/oxidase and triacylglycerol with glycerol kinase. LDL-cholesterol was calculated according to the Friedewald formula. Insulin was measured using radioimmunoassay (Invitrogen, Paisley, United Kingdom). HOMA-IR was calculated using fasting glucose and insulin concentrations (Matthews et al., 1985 (link)). The intra- and inter-assay coefficient of variation for all blood parameters was ≤5%.

Bowden Davies K.A., Norman J.A., Thompson A., Mitchell K.L., Harrold J.A., Halford J.C., Wilding J.P., Kemp G.J., Cuthbertson D.J, & Sprung V.S. (2021). Short-Term Physical Inactivity Induces Endothelial Dysfunction. Frontiers in Physiology, 12, 659834.

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Comprehensive Metabolic Assessment Protocol

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Anthropometric measurements Weight, height and waist and hip circumferences were measured. Participants then rested for 5 min before BP was determined from an average of three measures.
Biochemical measurements Blood samples were collected and analysed using the Olympus AU2700 analyser (Beckman Coulter, High Wycombe, UK) with standard proprietary reagents as follows: glucose with hexokinase, total cholesterol and HDL-cholesterol with cholesterol esterase/ oxidase and triacylglycerol with glycerol kinase. LDLcholesterol was calculated according to the Friedewald formula. Insulin was measured using radioimmunoassay (Invitrogen, Paisley, UK). HOMA-IR was calculated using fasting glucose and insulin concentrations [24] . NEFA was measured using a NEFA assay kit (Randox Daytona, County Antrim, UK). Fasting NEFA and insulin concentrations were used to estimate adipose tissue insulin resistance (adipo-IR) [25] .
OGTT After fasting blood samples were collected, a 75 g glucose solution was consumed within 5 min and post-ingestion blood samples were drawn at 30, 60, 90 and 120 min. Glucose, insulin and NEFA responses were calculated as AUC. Matsuda index was calculated to estimate whole-body insulin sensitivity; indices of hepatic insulin resistance and skeletal muscle insulin sensitivity were determined as previously described [26, 27] .

Bowden Davies K.A., Sprung V.S., Norman J.A., Thompson A., Mitchell K.L., Halford J.C.G., Harrold J.A., Wilding J.P.H., Kemp G.J, & Cuthbertson D.J. (2018). Short-term decreased physical activity with increased sedentary behaviour causes metabolic derangements and altered body composition: effects in individuals with and without a first-degree relative with type 2 diabetes. Diabetologia, 61(6).

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