Rat leptin elisa kit

Manufactured by Merck Group

Sourced in Spain, United States

The Rat Leptin ELISA Kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the measurement of leptin levels in rat samples.

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15 protocols using rat leptin elisa kit

Plasma Biomarker Profiling Protocol

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Alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) activities, low-density lipoprotein (LDL), high-density lipoprotein (HDL) and total cholesterol levels were determined in plasma samples by the Instrumental Techniques Laboratory of the University of León using standard techniques.
Insulin plasma concentration was determined using the commercial kit Rat/Mouse Insulin ELISA kit (EZRMI-13, Sigma-Aldrich; St. Luis, MO, USA), whereas leptin plasma concentration was quantified using Rat leptin ELISA kit (RAB0335, Sigma-Aldrich; St. Luis, MO, USA) according to manufacturer’s guidelines.

Juárez-Fernández M., Porras D., Petrov P., Román-Sagüillo S., García-Mediavilla M.V., Soluyanova P., Martínez-Flórez S., González-Gallego J., Nistal E., Jover R, & Sánchez-Campos S. (2021). The Synbiotic Combination of Akkermansia muciniphila and Quercetin Ameliorates Early Obesity and NAFLD through Gut Microbiota Reshaping and Bile Acid Metabolism Modulation. Antioxidants, 10(12), 2001.

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Rodent Metabolic Profiling Protocol

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The rats were fasted for 16 h, and blood was collected by tail vein puncture and the serum chemistry parameters were analyzed at Shanghai Changzheng Hospital, the Secondary Military Medical University.
For glucose tolerance test, the animals were fasted for 6 h before administered ip injection with d-glucose at 2 g/kg, and blood was collected by tail vein puncture. The glucose levels were measured by Accu-Chek Performa (Roche).
The animals were fasted for 16 h and blood was collected by tail vein puncture. Blood was left to clot for 1 h at room temperature, and serum was obtained after centrifugation. The serum INSULIN levels were measured by Rat/Mouse Insulin ELISA Kit 96-well plate (Millipore). The serum LEPTIN level was measured by Rat Leptin ELISA Kit (Sigma).

Xu S., Zhu X., Li H., Hu Y., Zhou J., He D., Feng Y., Lu L., Du G., Hu Y., Liu T., Wang Z., Ding G., Chen J., Gao S., Wu F., Xue Z., Li Y, & Fan G. (2016). The 14th Ile residue is essential for Leptin function in regulating energy homeostasis in rat. Scientific Reports, 6, 28508.

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Serum Biomarker Analysis Protocol

Cited 1 time

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Analysis of serum samples was performed as in [44 (link)]. Briefly, enzymatic colorimetric kits were used for the determination of serum total cholesterol, triglycerides, and glucose (QCA, Barcelona, Spain). Serum insulin and leptin levels were measured using a mouse/rat insulin ELISA kit (Mercodia, Barcelona, Spain) and a rat leptin ELISA kit (Millipore, Barcelona, Spain). The homeostasis model assessment-estimated insulin resistance (HOMA-IR) was calculated using the following formula [46 (link)]: HOMA-IR = glucose × insulin/22.5.

Alvarez-Monell A., Subias-Gusils A., Mariné-Casadó R., Boqué N., Caimari A., Solanas M, & Escorihuela R.M. (2022). Impact of Calorie-Restricted Cafeteria Diet and Treadmill Exercise on Sweet Taste in Diet-Induced Obese Female and Male Rats. Nutrients, 15(1), 144.

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Reproductive Hormone and Metabolite Analysis

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Sex hormones including follicle-stimulating hormone (FSH), luteinizing hormone (LH),
estradiol (E2), progesterone (P), testosterone (T), prolactin (PRL), and other
reproduction-related hormones including LEP, insulin-like growth factor-1 (IGF1),
cortisol, growth hormone (GH),and Ca, and Pi in the serum were determined by Nanjing
Jiancheng Bioengineering Institute (Nanjing, China). CCK levels were measured using a rat
CCK enzyme-linked immunosorbent assay (ELISA) kit (Groundwork Biotechnology Diagnosticate,
San Diego, CA, USA). Serum LEP levels were measured using a rat leptin ELISA kit
(Millipore Corporation, Billerica, MA, USA).

Wu L., Liu W., Bayaer N., Gu W, & Song J. (2014). Exogenous Leptin Administered Intramuscularly Induces Sex Hormone Disorder and Ca Loss via Downregulation of Gnrh and PI3K Expression. Experimental Animals, 63(4), 447-457.

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Circulating Metabolic Biomarker Profiling

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Enzymatic colorimetric kits were used to determine the circulating levels of triacylglycerols, glucose, total cholesterol (QCA, Barcelona, Spain), phospholipids (phosphatidylcholine) (Spinreact, Girona, Spain) and non-esterified free fatty acids (FFAs) (WAKO, Neuss, Germany). The plasma levels of insulin, leptin, monocyte chemoattractant protein-1 (MCP-1), and adiponectin were measured using a rat insulin ELISA kit (Mercodia, Upssala, Sweden), a rat leptin ELISA kit (Millipore, Barcelona, Spain), a rat MCP-1 ELISA kit (Thermo Scientific, Rockford, IL, USA), and a rat adiponectin ELISA kit (Millipore, Barcelona, Spain), respectively.

Mariné-Casadó R., Domenech-Coca C., Crescenti A., Rodríguez Gómez M.Á., Del Bas J.M., Arola L., Boqué N, & Caimari A. (2022). Maternal Supplementation with a Cocoa Extract during Lactation Deeply Modulates Dams’ Metabolism, Increases Adiponectin Circulating Levels and Improves the Inflammatory Profile in Obese Rat Offspring. Nutrients, 14(23), 5134.

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Enzymatic Colorimetric Assay of Serum Lipids

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An enzymatic colorimetric kit was used to assay serum triacylglycerides (QCA, Barcelona, Spain). Serum insulin and leptin levels were measured using a mouse/rat insulin ELISA kit (Millipore, Barcelona, Spain) and a rat leptin ELISA kit (Millipore), respectively. Insulin resistance was estimated using the homeostatic model assessment (HOMA-IR), following the formula: HOMA-IR = Glucose × Insulin/22.5 [28 (link)].

Subias-Gusils A., Álvarez-Monell A., Boqué N., Caimari A., Del Bas J.M., Mariné-Casadó R., Solanas M, & Escorihuela R.M. (2021). Behavioral and Metabolic Effects of a Calorie-Restricted Cafeteria Diet and Oleuropein Supplementation in Obese Male Rats. Nutrients, 13(12), 4474.

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Serum Biomarker Quantification Protocol

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Enzymatic colorimetric kits were used for the determination of serum glucose, triglyceride, cholesterol (QCA, Barcelona, Spain) and non-esterified free fatty acid (NEFA) (WAKO, Neuss, Germany) levels. The serum insulin and leptin levels were measured using a mouse/rat insulin ELISA kit and a rat leptin ELISA kit, respectively, (Millipore, Barcelona, Spain) following the manufacturer’s instructions.

Cigarroa I., Lalanza J.F., Caimari A., del Bas J.M., Capdevila L., Arola L, & Escorihuela R.M. (2016). Treadmill Intervention Attenuates the Cafeteria Diet-Induced Impairment of Stress-Coping Strategies in Young Adult Female Rats. PLoS ONE, 11(4), e0153687.

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Serum IGF-1 and Leptin Quantification

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Serum was separated by centrifugation at 450 g for 10 min at 4 °C and stored at −70 °C. The chemical analysis of the serum was performed by American Medical Laboratories (AML) Ltd. (Herzlia, Israel). Serum levels of insulin-like growth factor-1 (IGF-1) and leptin were measured using commercial kits according to the manufacturer’s recommendations: Quantikine Mouse/Rat IGF-1 assay kit, detection limit 8.4 pg/mL (Cat. No. MG100, R & D Systems, Minneapolis, MN, USA); Rat Leptin ELISA kit, detection limit 22 pg/mL (Millipore, Billerica, MA, USA).

Bar-Maisels M., Gabet Y., Shamir R., Hiram-Bab S., Pasmanik-Chor M., Phillip M., Bar-Yoseph F, & Gat-Yablonski G. (2017). Beta Palmitate Improves Bone Length and Quality during Catch-Up Growth in Young Rats. Nutrients, 9(7), 764.

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Biochemical Assays for Metabolic Profile

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Enzymatic colorimetric kits were used for the determination of serum total cholesterol, triglycerides, and glucose (QCA, Barcelona, Spain), HDL-cholesterol and LDL-cholesterol (Bioassay systems, CA, USA) and non-esterified free fatty acids (NEFAs) (WAKO, Neuss, Germany). Serum insulin and leptin levels were measured using a mouse/rat insulin ELISA kit (Millipore, Barcelona, Spain) and a rat leptin ELISA kit (Millipore). The homeostasis model assessment-estimated insulin resistance (HOMA-IR) was calculated following the formula^{40 (link)}: HOMA-IR = Glucose(mM) X Insulin(mU/L)/22.5.

Alvarez-Monell A., Subias-Gusils A., Mariné-Casadó R., Belda X., Gagliano H., Pozo O.J., Boqué N., Caimari A., Armario A., Solanas M, & Escorihuela R.M. (2022). Restricted cafeteria feeding and treadmill exercise improved body composition, metabolic profile and exploratory behavior in obese male rats. Scientific Reports, 12, 19545.

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Serum Biomarkers in Rat Studies

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All rats were euthanised by CO 2 inhalation at the end of the experimental period and blood was collected by cardiac puncture. Serum was separated by centrifugation at 1500 rpm (239 g) in a Rotina 46 R centrifuge (Hettich) for 10 min at 4°C and stored at -70°C. For the short-term experiment only, serum levels of total Ca, P and total alkaline phosphatase (ALP) were measured using a Beckman Coulter analyzer (AU 680; Beckman Coulter International). For both the short-and long-term experiments, serum levels of insulin-like growth factor-I (IGF-I), leptin and osteocalcin (OC, bone γ-carboxyglutamic acidcontaining protein) were determined using commercial kits according to the manufacturer's recommendations as follows: Quantikine Mouse/Rat IGF-I assay kit, detection limit 8•4 pg/ml (cat. no. MG100; R&D Systems); Rat Leptin ELISA kit, detection limit 22 pg/ml (Millipore); and rat osteocalcin (OC) ELISA assay kit, detection limit 0•5 ng/ml (cat. no. EIA-2095; DRG International Inc.).

Masarwi M., Gabet Y., Dolkart O., Brosh T., Shamir R., Phillip M, & Gat-Yablonski G. (2016). Skeletal effect of casein and whey protein intake during catch-up growth in young male Sprague-Dawley rats. The British journal of nutrition, 116(1).

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