This Human LEP ELISA Kit was designed for the quantitative measurement of Human LEP protein in serum, plasma, tissue homogenates. It is a Sandwich ELISA kit, its detection range is 0.156 ng/mL-10 ng/mL and the sensitivity is 0.060 ng/mL.

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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. 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. 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. 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. May also play an apoptotic role via JAK2-STAT3 pathway and up-regulation of BIRC5 expression. 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). 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. 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. In adaptive immunity, promotes the switch of memory T-cells towards T helper-1 cell immune responses. Increases CD4(+)CD25(-) T-cell proliferation and reduces autophagy during TCR (T-cell receptor) stimulation, through MTOR signaling pathway activation and BCL2 up-regulation.

• A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human LEP antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
• Two vials lyophilized standard --- Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
• One vial Biotin-labeled LEP antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
• One vial HRP-avidin (100 x concentrate) (120 μl/bottle) --- Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
• One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody solution.
• One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
• One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
• One vial Wash Buffer (25 x concentrate) (20 ml/bottle) --- Wash away unbound or free substances.
• One vial TMB Substrate (10 ml/bottle) --- Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
• One vial Stop Solution (10 ml/bottle) --- Stop the color reaction. The solution color immediately turns from blue to yellow.
• Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
• An instruction manual

• A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
• An incubator can provide stable incubation conditions up to 37°C±5°C.
• Centrifuge
• Vortex
• Squirt bottle, manifold dispenser, or automated microplate washer
• Absorbent paper for blotting the microtiter plate
• 50-300ul multi-channel micropipette
• Pipette tips
• Single-channel micropipette with different ranges
• 100ml and 500ml graduated cylinders
• Deionized or distilled water
• Timer
• Test tubes for dilution