Mouse Leptin ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Mouse Leptin with a sensitivity of 21.4 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Application | Reactivity | Dilution info | Notes |
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Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Key player in the regulation of energy balance and body weight control. Once released into the circulation, has central and peripheral effects by binding LEPR, found in many tissues, which results in the activation of several major signaling pathways (PubMed:11373681, PubMed:12594516, PubMed:15899045, PubMed:16825198, PubMed:20620997). In the hypothalamus, acts as an appetite-regulating factor that induces a decrease in food intake and an increase in energy consumption by inducing anorexinogenic factors and suppressing orexigenic neuropeptides, also regulates bone mass and secretion of hypothalamo-pituitary-adrenal hormones. In the periphery, increases basal metabolism, influences reproductive function, regulates pancreatic beta-cell function and insulin secretion, is pro-angiogenic for endothelial cell and affects innate and adaptive immunity (By similarity) (PubMed:10660043, PubMed:12594516, PubMed:25060689, PubMed:25383904, PubMed:8589726, PubMed:9732873). In the arcuate nucleus of the hypothalamus, activates by depolarization POMC neurons inducing FOS and SOCS3 expression to release anorexigenic peptides and inhibits by hyperpolarization NPY neurons inducing SOCS3 with a consequent reduction on release of orexigenic peptides (By similarity) (PubMed:11373681, PubMed:20620997). In addition to its known satiety inducing effect, has a modulatory role in nutrient absorption. In the intestine, reduces glucose absorption by enterocytes by activating PKC and leading to a sequential activation of p38, PI3K and ERK signaling pathways which exerts an inhibitory effect on glucose absorption. Acts as a growth factor on certain tissues, through the activation of different signaling pathways increases expression of genes involved in cell cycle regulation such as CCND1, via JAK2-STAT3 pathway, or VEGFA, via MAPK1/3 and PI3K-AKT1 pathways (By similarity) (PubMed:16825198, PubMed:20620997). May also play an apoptotic role via JAK2-STAT3 pathway and up-regulation of BIRC5 expression (By similarity). Pro-angiogenic, has mitogenic activity on vascular endothelial cells and plays a role in matrix remodeling by regulating the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) (PubMed:16825198). In innate immunity, modulates the activity and function of neutrophils by increasing chemotaxis and the secretion of oxygen radicals. Increases phagocytosis by macrophages and enhances secretion of pro-inflammatory mediators. Increases cytotoxic ability of NK cells (Probable). Plays a pro-inflammatory role, in synergy with IL1B, by inducing NOS2 wich promotes the production of IL6, IL8 and Prostaglandin E2, through a signaling pathway that involves JAK2, PI3K, MAP2K1/MEK1 and MAPK14/p38 (PubMed:15899045). In adaptive immunity, promotes the switch of memory T-cells towards T helper-1 cell immune responses (By similarity). Increases CD4(+)CD25(-) T cells proliferation and reduces autophagy during TCR (T cell receptor) stimulation, through MTOR signaling pathway activation and BCL2 up-regulation (PubMed:25060689).
Ob, Lep, Leptin, Obesity factor
Mouse Leptin ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Mouse Leptin with a sensitivity of 21.4 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Sample | n | C.V. |
---|---|---|
Sample Mouse Serum | n 5 | C.V. 5.9 |
Sample | n | C.V. |
---|---|---|
Sample Mouse Serum | n 3 | C.V. 11.5 |
Sample type | Average % | Range |
---|---|---|
Sample type Serum | Average % = 111 | Range 110 - 114 % |
Sample type Urine | Average % = 80 | Range 77 - 82 % |
Sample type Heparin Plasma | Average % = 106 | Range 94 - 116 % |
Sample type Citrate plasma | Average % = 104 | Range 100 - 111 % |
Sample type Cell culture media | Average % = 104 | Range 103 - 105 % |
Leptin in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Leptin protein in mouse serum, plasma (citrate), urine and cell culture supernatants.
This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
If using a Molecular Devices' plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.
The CatchPoint SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.
Leptin is a secreted protein factor produced by adipocytes. It regulates energy balance and body fat deposits. Leptin deficiency in humans and mice can cause obesity. Circulating levels of Leptin are regulated by food intake, insulin levels and pregnancy status. Mouse leptin has 96% and 85% protein sequence identity to rat and human leptin, respectively.
Leptin also known as the OB protein is a hormone with a molecular weight of approximately 16 kDa. Primarily it is produced by adipocytes in white adipose tissue and to a lesser extent in the placenta stomach and other tissues. This protein plays a significant role in energy homeostasis by conveying information to the central nervous system regarding fat storage. In scientific studies leptin is often measured using methods like the leptin ELISA kit or mouse leptin assay to monitor its levels and function.
Leptin regulates food intake and energy expenditure by binding to its receptors in the hypothalamus. It is an important part of the feedback loop to manage body weight and appetite. Leptin is not part of a larger protein complex and acts directly on its target tissues. By signal transduction pathways it influences cellular processes such as metabolism and even impacts fertility and immune responses.
Leptin is involved in the neuroendocrine signaling pathway and the MAPK/ERK pathway. In the neuroendocrine system leptin interacts with proteins like proopiomelanocortin (POMC) and neuropeptide Y (NPY) to manage appetite and energy use. Additionally it activates the JAK/STAT pathway when binding to its receptor which influences gene expression involved in metabolism and inflammation.
Leptin is linked to obesity and type 2 diabetes. Leptin deficiencies or resistance can lead to metabolic disorders as the hormone's signaling is disrupted affecting energy balance and storage. Its connection to adiponectin another hormone derived from fat tissue is critical in understanding these conditions; while leptin promotes inflammation adiponectin has anti-inflammatory effects. Exploring these proteins' roles and their balance offers insights into treatments for obesity-related disorders.
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Example of mouse Leptin standard curve in Sample Diluent NS.
Background-subtracted data values (mean +/- SD) are graphed.
Interpolated concentrations of Leptin in mouse serum and plasma (citrate).
The concentrations of Leptin were measured in duplicate and interpolated from the Leptin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Leptin concentration was determined to be 4514.3 pg/mL in mouse serum and 2698.1 pg/mL in mouse plasma (Citrate).
Interpolated concentrations of Leptin in rat serum, plasma (citrate), and plasma (heparin).
The concentrations of Leptin were measured in duplicate and interpolated from the Leptin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Leptin concentration was determined to be 2983 pg/mL in rat serum, 1626 pg/mL in rat plasma (citrate) and 2134 pg/mL in rat plasma (heparin).
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