LEP ELISA Kit (Sheep)

Katalog-Nummer OKCA01024

Size : 96Wells

Marke : Aviva Systems Biology

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Datasheets/ManualsClick here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
Product Info
Predicted Species ReactivitySheep
ELISA Kit Detection MethodColorimetric, OD450 nm
ELISA Kit Duration~ 3.5 Hours
ELISA Kit Linearity
DilutionMean n=4 (%)Range (%)
Neat9082-98
1:29386-100
1:49588-102
1:89590-100
ELISA Kit PrincipleAviva Systems Biology LEP ELISA Kit (Sheep) (OKCA01024) is based on standard sandwich enzyme-linked immuno-sorbent assay technology. A antibody specific for LEP has been pre-coated onto a 96-wellplate (12 x 8 Well Strips). Standards or test samples are added to the wells, incubated and removed. A biotinylated detector antibody specific for LEP is added, incubated and followed by washing. Avidin-Peroxidase Conjugate is then added, incubated and unbound conjugate is washed away. An enzymatic reaction is produced through the addition of TMB substrate which is catalyzed by HRP to generating a blue color product that changes yellow after adding acidicstop solution. The density of yellow coloration read by absorbance at 450 nm and is quantitatively proportional to the amount of sample LEP captured in well.
ELISA Kit Range3.12 ng/mL-200 ng/mL
ELISA Kit Recovery
MatrixRecovery range (%)Average(%)
Serum (n=5)9288-96
EDTA Plasma (n=4)9590-101
ELISA Kit ReproducibilityIntra-assay Precision: 3 samples with known low, middle and high level LEP were tested with 20 replicates across one plate.
Inter-assay Precision: 3 samples with known low, middle and high level LEP were tested on three different plates with 8 replicates, respectively.
Intra-Assay CV: <8%
Inter-Assay CV: <10%
ELISA Kit Component
ComponentAmount
LEP Microplate96 Wells (12 x 8 Well strips)
LEP Lyophilized Standard2
100X Biotinylated LEP Detector Anitbody1 x 120 uL
100X Avidin-HRP Conjugate1 x 120 uL
Sample Diluent1 x 50 mL
Detector Antibody Diluent1 x 15 mL
Conjugate Diluent1 x 15 mL
25X Wash Buffer1 x 20 mL
Stop Solution1 x 10 mL
TMB Substrate1 x 10 mL
Reconstitution and StorageStore as indicated in product manual.
Sample TypeSerum, Plasma, Tissue Homogenates
Sensitivity0.78 ng/mL
Assay InfoAssay Methodology: Quantitative Sandwich ELISA
Protocol Information
  • Reconstitution & Storage Instructions
  • Western Blotting/Immunoblotting (WB/IB) Protocol
  • Immunohistochemistry (IHC) Protocol
  • Immunocytochemistry (ICC) Protocol
  • Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol
  • Blocking Peptide Competition Protocol (BPCP)
  • Immunoprecipitation (IP) Protocol
  • Antibody Array (AA) Protocol
Gene SymbolLEP
Gene Full NameLeptin
Protein NameLeptin
Description of TargetKey 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.
Uniprot IDQ28603
Protein Accession #XP_004008087.2
Nucleotide Accession #XM_004008038.3