Leptin concentrations in plasma were measured with the enzyme-linked immunosorbent assay, using the Human Leptin ELISA kit (Millipore; sensitivity 0.5 ng/mL). The absorbance was measured at 450/630 nm using the STAR FAX 303/PLUS (Awareness Technology, Inc.). Leptin concentrations were measured in triplicate and calculated from standard curves generated for each assay from the recombinant human leptin provided in the kit.
Human leptin elisa kit
Manufactured by Merck Group
Sourced in United States, Germany
The Human Leptin ELISA kit is a quantitative sandwich enzyme immunoassay designed for the measurement of leptin in human serum, plasma, and other biological fluids. Leptin is a hormone produced by adipose tissue that regulates energy balance and body weight.
Automatically generated - may contain errors
Lab products found in correlation
Duoset elisa development kit, by R&D Systems (1 mentions)
Spectramax 190, by Molecular Devices (1 mentions)
7600 autoanalyzer, by Hitachi (1 mentions)
Lipolysis assay kit, by Abcam (1 mentions)
Xn 10, by Sysmex (1 mentions)
Human il 6 quantikine hs elisa, by R&D Systems (1 mentions)
Human tnf α quantikine hs elisa, by R&D Systems (1 mentions)
12 protocols using human leptin elisa kit
1
Leptin Quantification in Plasma
2
Serum Leptin Levels After Fasting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Serum leptin levels were measured after an overnight fast. Overnight fasting blood sampling was undertaken by a medical laboratory technologist. After collection of blood samples, the serum was transferred to cryotubes and stored in a freezer at −80°C. All serum parameters were analyzed by enzyme-linked immunosorbent assay using commercially available kits (Human Leptin ELISA kit, Millipore, Billerica, MA, United States) at the professional clinical laboratory services unit of a biotechnical company (Green Cross Reference Lab, Seoul, South Korea).
3
Hormonal Biomarker Quantification Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Leptin, ghrelin, GLP-1 and PYY levels were determined with commercially available reagent kits from Millipore, St. Charles, U.S. Leptin serum levels were determined with the immunoenzymatic technique ELISA using the Human Leptin ELISA Kit. The method was characterised by sensitivity of 0.78 ng/ml, and inter- and intra-assay coefficients of variability below 4.6% and 6.2%, respectively.
Ghrelin levels were determined with the immunoenzymatic technique using the Human Ghrelin (Total) ELISA Kit. The method determined two ghrelin forms: active and des-octanoyl. The method sensitivity limit corresponded to 0.50 pg/ml, and inter- and intra-assay coefficients of variability were below 1.91% and 7.81%, respectively. The serum levels of active GLP-1 were determined with the immunoenzymatic technique based on fluorescence detection, using fluorogenic substrate for alkaline phosphatase, 4-methylumbelliferyl phosphate. The Glucagon-Like Peptide-1 (Active) ELISA Kit used was characterised by sensitivity of 2 pM and inter- and intra-assay coefficients of variability below 9% and 13%, respectively. Peptide YY levels were determined with the immunoenzymatic technique using the Human PYY (Total) ELISA Kit. The analytical sensitivity of this method was 16.07 pg/ml, while inter- and intra-assay coefficients of variability did not exceed 5.78% and 16.50%, respectively.
Ghrelin levels were determined with the immunoenzymatic technique using the Human Ghrelin (Total) ELISA Kit. The method determined two ghrelin forms: active and des-octanoyl. The method sensitivity limit corresponded to 0.50 pg/ml, and inter- and intra-assay coefficients of variability were below 1.91% and 7.81%, respectively. The serum levels of active GLP-1 were determined with the immunoenzymatic technique based on fluorescence detection, using fluorogenic substrate for alkaline phosphatase, 4-methylumbelliferyl phosphate. The Glucagon-Like Peptide-1 (Active) ELISA Kit used was characterised by sensitivity of 2 pM and inter- and intra-assay coefficients of variability below 9% and 13%, respectively. Peptide YY levels were determined with the immunoenzymatic technique using the Human PYY (Total) ELISA Kit. The analytical sensitivity of this method was 16.07 pg/ml, while inter- and intra-assay coefficients of variability did not exceed 5.78% and 16.50%, respectively.
4
Biomarkers of Insulin Resistance in Fasting Blood Samples
5
Quantification of Adipocytokines Using ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Commercially available ELISA kits were used to quantify adipocytokines, in accordance with the manufacturer's instructions (Millipore, Human Leptin ELISA kit, Cat. # EZHL-80SK; Human High Molecular Weight Adiponectin ELISA kit, Cat. # EZHMWA-64K; Human Resistin ELISA kit, Cat. # EZHR-95K). The intra- and interassay coefficients of variation for leptin, adiponectin, and resistin were 3.5% and 6.5%, 3.2% and 4.4%, and 4.7% and 8.4%, respectively. When calculating adiponectin/leptin and adiponectin/resistin ratios, adiponectin values were converted from μg/mL to ng/mL.
6
Blood Panel and Adipokine Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Details of the sample collection were described in a previous study [7 (link)]. After a fasting period of 12 h, venous blood specimens were collected and stored at − 70 °C until use in further analysis. Levels of the lipid panel, free fatty acids, a glucose-related panel, serum gamma-glutamyltransferase (gamma-GT) assayed via a modified Sanz method, and serum high-sensitivity C-reactive protein (hs-CRP) assayed via a latex-agglutination turbidimetric immunoassay were measured by a Hitachi 7600 autoanalyzer (Hitachi, Tokyo, Japan). Plasma adiponectin concentrations were measured by an enzyme immunoassay [11 (link)], and leptin was measured with a Human Leptin ELISA kit (Millipore, Darmstadt, Germany) by SpectraMax190 (Molecular Devices, Shanghai, China).
7
Metabolic Hormone Analysis in Fasting Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Venous samples were collected (10 ml) in the mornings while fasting, at all three different follow-up times. The concentrations of insulin (IU/ml) and cortisol (µg/dl) were analyzed by means of chemiluminescence (active insulin Enzyme-Linked Immunosorbent Assay [ELISA] and assay design cortisol ELISA kits, Labtest, Lagoa Santa, Minas Gerais, Brazil). Ghrelin levels (pg/ml) were analyzed using a total human ghrelin ELISA kit (Sigma-Aldrich, St Louis, Missouri, USA). Leptin levels (ng/ml) were measured using a human leptin ELISA kit (Sigma-Aldrich, St Louis, Missouri, USA). All analyses were carried out in accordance with the manufacturer’s recommended instructions.
8
Adipose-Derived Stem Cell Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Adipose derived mesenchymal stem cells were obtained from Himedia TM Cat.# CCK011. Complete medium and adipogeneic differentiation medium were also purchased from Himedia Cat.# AL521 and Cat.# AL537 respectively. Cytotoxicity kit was purchased from Promega TM Cat.# G1780 to check the cell viability. Free glycerol and Adiponectin were measured using Abcam's Lipolysis Assay kit (Cat. #ab185433) and Human Adiponectin ELISA kit (Cat.#ab99968). Other adipocytokines were quantified using the kits from Sigma Aldrich: Human Leptin Elisa kit (RAB0333), Human Interleukin-8(IL-8) kit (Cat. # RAB0319) and Human resistin ELISA kit (Cat. # RAb0419).
9
Serum Adiponectin and Leptin Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Adiponectin and leptin levels were measured in all collected blood samples. Blood samples were immediately processed after collection and serum samples were stored at −80 °C till analysis. Serum samples were thawed on ice, diluted, and measured with commercially available ELISA kits (Human ADIPQ ELISA kit and Human Leptin ELISA kit, Merck life Sciences, Bengaluru, India). Assays were performed according to the provided protocol with 10% of the samples measured in duplicate. A linear regression standard curve was generated using GraphPad Prism (version 5) to interpolate sample values.
10
Venous Blood Sampling and Biobanking
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Venous blood samples were collected at least 2 h postprandially. Local anesthetic topical cream was used prior to sampling. The tubes were centrifuged for 10 min at 1300× g. Serum was separated into Cryo tubes and then stored at −80 °C at the Biobank in Lund or Umeå until analysis. Hemoglobin (Hb) was analyzed directly at the University and Regional Laboratories of Skåne or Norrland’s University Hospital with the Sysmex XN-10 (Sysmex Corporation, Chuoko, Japan). The tubes were transported from the Biobanks frozen on dry ice to the respective laboratory for analysis. Serum (s) insulin, C-peptide, and IGF-1 were analyzed at the University and Regional Laboratories of Skåne, insulin and C-peptide with the Cobas 601 instrument (Roche Diagnostics, Rotkreuz, Switzerland) and IGF-1 by the IDS-iSYS assay (Immunodiagnostic System Ltd., Boldon, Tyne & Wear, UK). Serum leptin and leptin receptor were analyzed by ELISA (Human Leptin ELISA kit, EMD Millipore; Merck KGaA, Darmstadt, Germany, and Human Leptin R Quantikine® ELISA, R&D Systems Inc., Minneapolis, MN, USA) at the Pediatric Research Laboratory at Umeå University.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!