Blood samples were obtained just before the start of the dialysis session. Levels of serum albumin, calcium, and phosphate were measured by using routine laboratory methods. Aliquots of serum were stored at −20 °C while waiting for subsequent assay, and measurements were made immediately after thawing. The biochemical parameters of calcium metabolism were determined as previously described26 (link), 27 (link). Serum intact PTH levels were measured by using an electrochemiluminescence immunoassay kit (Elecsys PTH; Roche Diagnostics GmbH, Mannheim, Germany). Serum hs-CRP was measured through a nephelometric assay (SRL Inc., Tokyo Japan). Serum adiponectin levels were measured by using a human adiponectin ELISA kit (Otsuka Pharmaceuticals Co., Tokyo, Japan)1 (link). Serum leptin levels were measured with a human leptin RIA kit (Millipore Co., Billerica, MA, USA)28 (link).
Human leptin ria kit
Manufactured by Merck Group
Sourced in Japan
The Merck Human Leptin RIA Kit is a radioimmunoassay (RIA) used for the quantitative determination of leptin levels in human serum, plasma, or other biological samples. Leptin is a hormone produced by adipose tissue that plays a role in regulating energy balance and body weight.
Automatically generated - may contain errors
Lab products found in correlation
Cica liquid glu j, by Kanto Chemical (3 mentions)
Lumipulse presto insulin, by Fujirebio (3 mentions)
Rapidia auto hba1c l, by Fujirebio (3 mentions)
Elecsys pth, by Roche (1 mentions)
Human leptin quantikine, by R&D Systems (1 mentions)
Quantikine hs elisa, by R&D Systems (1 mentions)
Cobas modular system, by Roche (1 mentions)
Human adiponectin elisa kit, by Merck Group (1 mentions)
Human adipocyte fabp elisa kit, by BioVendor (1 mentions)
Luminex 200 instrument, by Merck Group (1 mentions)
7 protocols using human leptin ria kit
1
Serum Biomarker Analysis in Dialysis Patients
2
Assaying Ghrelin and Leptin Levels
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The concentrations of des-acyl ghrelin and active ghrelin in the plasma were assayed using a Desacyl-Ghrelin ELISA Kit (SCETI, Tokyo, Japan) and Active Ghrelin ELISA Kit (SCETI), respectively. The specific antibodies used in the former and latter assay kits recognise the C-terminal portion and the octanoyl-modified Ser 3 residue of ghrelin, respectively.10 (link) The concentration of leptin in the serum was assayed using Human Leptin RIA Kit (Millipore, Belmopan, Belize).
3
Comprehensive Metabolic Biomarker Profiling
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HbA1c, glucose, insulin, leptin, adiponectin, RBP4, glycoalbumin, and high sensitivity-C-reactive protein (hs-CRP) were estimated using commercially available ELISA kits (HbA1c; RAPIDIA Auto HbA1c-L [Fujirebio Inc. (Tokyo, Japan)]; glucose, Cica liquid GLU J [KANTO CHEMICAL Co., Inc. (Tokyo, Japan)]; insulin, Lumipulse Presto Insulin [Fujirebio Inc. (Tokyo, Japan)]; leptin, Human leptin RIA kit [Millipore Co., Inc. (Tokyo, Japan)]; adiponectin, CircuLex Human adiponectin ELISA Kit CY-8050 [MBL Co. Ltd. (Nagano, Japan)]; RBP4, CircuLex Human RBP4 ELISA Kit [MBL Co. Ltd. (Nagano, Japan)]; glycoalbumin, Lucia GA-L [Asahi KASEI Pharma Co., Inc. (Tokyo, Japan)]; and hs-CRP, CircuLex Human HS-CRP ELISA Kit CY-8071 [MBL Co. Ltd. (Nagano, Japan)]).
4
Comprehensive Metabolic and Biomarker Assessment
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HbA1c, glucose, insulin, leptin, adiponectin, RBP4, glycoalbumin, and hs-CRP were measured using commercially available ELISA kits {HbA1c: RAPIDIA Auto HbA1c-L [Fujirebio Inc. (Tokyo, Japan)]; glucose: Cica liquid GLU J [KANTO CHEMICAL Co., Inc. (Tokyo, Japan)]; insulin: Lumipulse Presto Insulin (Fujirebio Inc.); leptin: Human leptin RIA kit [Millipore Co., Inc. (Tokyo, Japan)]; adiponectin: CircuLexTM Human adiponectin ELISA Kit CY-8050 [MBL Co., Ltd. (Nagano, Japan)]; RBP4: CircuLex Human RBP4 ELISA Kit (MBL Co., Ltd.); glycoalbumin: Lucia GA-L [Asahi KASEI Pharma Co., Inc. (Tokyo, Japan)]; and hs-CRP: CircuLexTM Human HS-CRP ELISA Kit CY-8071 (MBL Co., Ltd.)}. Oxidative stress markers were measured according to our previous reports.(7 (link),8 (link),19 (link)) In this study, normal HbA1c was defined at 4.6 to 5.5%, while HbA1c levels >6.5% were considered to indicate diabetes. The HbA1c levels of 5.5–6.5% are considered to be borderline, and the diagnosis and treatment by physicians vary depending on the the protocol followed in the clinical setting in different countries.
5
Glycemic and Inflammatory Biomarker Levels
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Plasma levels of glycated hemoglobin (HbA1c), glucose, insulin, leptin, adiponectin, RBP4, glycated albumin, and high-sensitivity C-reactive protein (hs-CRP) were measured using the following commercially available enzyme-linked immunosorbent assay (ELISA) kits: HbA1c, RAPIDIA Auto HbA1c-L (Fujirebio, Tokyo, Japan); glucose, Cica Liquid GLU J (Kanto Chemical Co., Tokyo, Japan); insulin, Lumipulse Presto Insulin (Fujirebio, Tokyo, Japan); leptin, Human Leptin RIA kit (Millipore, Tokyo, Japan); adiponectin, CircuLex Human Adiponectin ELISA kit CY-8050 (MBL Co., Nagano, Japan); RBP4, CircuLex Human RBP4 ELISA kit (MBL Co.); glycated albumin, Lucia GA-L (Asahi KASEI Pharma Co., Tokyo, Japan); and hs-CRP, CircuLex Human HS-CRP ELISA kit CY-8071 (MBL Co.).
6
Maternal Adipokines and Gestational Diabetes
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All blood and medium samples were analyzed at the Clinical Chemistry Laboratory, Sahlgrenska University Hospital, accredited in accordance with the International Standard ISO 15189:2007. Enzyme-linked immunosorbent assay (ELISA) was used to measure adiponectin (Human adiponectin ELISA kit, Millipore, Billerica, MA, USA), tumor necrosis factor (TNF)-α (human TNF-alpha Quantikine HS ELISA, R&D Systems, Minneapolis, MN, USA) and adipocyte fatty acid-binding protein (AFABP) (Human Adipocyte FABP ELISA kit, BioVendor Research and Diagnostic Products, Bratislava, Slovakia). As a result of a change in assay during the study period, leptin was measured by RIA or ELISA (Human Leptin RIA kit, Millipore; Quantikine Human Leptin, R&D Systems; R=0.962). Samples from each woman in T1 and T3 were analyzed with the same leptin assay. HbA1c was measured by Mono-S ion-exchange chromatography. Glycerol was analyzed with a Randox radiometric glycerol kit (GY105; Crumlin, UK) and a Konelab 30 autoanalyzer (Thermo Clinical Labsystems, Vantaa, Finland). All other assays were routinely performed with a Cobas Modular system (Roche Diagnostics, Mannheim, Germany).
7
Cytokine and Leptin Quantification in Serum and Adipose Tissue
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Human cytokine quantification was performed using a Milliplex MAP kit for Human Cytokine/Chemokine magnetic bead panel, which measured five analytes in serum from nine paired patients before and 3 months after VSG. Six of the nine patients from this cohort also obtained a fat biopsy for mRNA analysis of adipose tissue. Serum samples were obtained in the morning after an overnight fast. Analytes included in the kit were IL‐6, IL‐10, IL‐β, and TNF‐α. All samples were acquired on a Luminex 200 instrument (Millipore) and were performed in duplicate in 96‐well plates. The serum concentration of leptin was determined by radioimmunoassay using the human leptin RIA kit (Millipore). The absorbance values were used to calculate the leptin concentration and were carried out in duplicate, and the average of the two values was used for data analysis.
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