More info
Assay range | 0.375-24 ng/ml |
Sensitivity | 0.30 ng/ml |
Specificity | No cross-reaction with other related substances detected |
Size | 96T |
Storage | Store at 2 - 8ºC. Keep reconstituted standard and detection Ab at -20 ºC |
Assay Principle | Sandwich ELISA |
Sample Volume | 50 µL final volume, dilution factor varies on samples |
Sample Type | plasma, serum, and cell culture supernatants |
Detection Method | Chromogenic |
Kit Components
1. Recombinant Mouse Leptin standard: 1 vial
2. One 96-well plate coated with Mouse Leptin Ab
3. Diluent buffer (10x): 30 mL - 1
4. Biotinylated Mouse Leptin Ab (50x) 110 µL
5. Streptavidin-HRP (100x): 80 µL
6. TMB developing agent: 8 mL x1
7. Stop solution: 12 mL x1
8. Washing solution (20x): 30 mL x2
Background
Leptin, encoded by ob/lep gene in humans, is a 146 amino acid (aa) residue non-glycosylated polypeptide. Human Leptin is 87% and 84% aa identical to mouse and rat Leptin, respectively, and is active in both the mouse and rat systems. Leptin is primarily expressed by adipocytes and its production is influenced by hormones, cytokines and nutrients. Leptin circulates in the plasma, crosses the blood brain barrier, and is present in human breast milk.
The human Leptin receptor (ObR or LEPR) is an 1144 aa residue type I transmembrane glycoprotein of the IL-6 receptor family. Several alternative spliced forms (a-f) of obR gene have been identified. ObRa, ObRc, ObRd and ObRf with truncated cytoplasmic domains, are responsible for mediating Leptin binding and endocytosis, but not signal transduction, while ObRb (formerly OB RL) containing a large cytoplasmic domains, expressed mainly in the hypothalamic arcuate nucleus, is essential for signal transduction. Mutations of ObRb can cause obese phenotypes in both the mouse and rat. In addition, a soluble Leptin R has been identified and is the primary Leptin-binding protein in blood to maintain a pool of available bioactive Leptin, delay Leptin clearance from circulation, and down-regulate blood brain transmission of Leptin. In a concentration-dependent manner, Leptin signaling leads to pro-opiomelanocortin (POMC) expressing neurons to reduce food intake, and neuropeptide Y (NpY) and agouti-related protein (AgRP) expressing neurons to increase food intake. Leptin concentrations are higher in women than in men. In women, serum leptin was the most important indicator of myocardial infarction. In patients with angiographically confirmed coronary atherosclerosis, leptin is a novel biomarker of future cardiovascular events independent of other risk factors, including lipid status and CRP.