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Manual Download| Product Name | FS-Mouse LEP(Leptin) ELISA Kit | Species | Mouse |
| Uniprot ID | P41160 | Alternative Names | FLJ94114, LEP, leptin (murine obesity homolog), leptin (obesity homolog, mouse), Leptin, OB, Obese protein, obese, mouse, homolog of, Obesity factor, OBOBS |
| Detection method | Sandwich | Sensitivity | 29.92 pg/mL |
| Standard | 5000pg/mL | Detection Range | 78.13-5000pg/mL |
| Sample type | Serum, Plasma, Tissue homogenates, Cell lysates, Cell culture supernates and Other biological fluids;Sample Volume=50μL | ||
| Reaction time | 1.5H | Research Area | Neuroscience, Signaling transduction, Cancer; |
| Test principle | This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse LEP. Samples (or Standards) and Horseradish Peroxidase (HRP) linked antibody specific for Mouse LEP are added to the micro ELISA plate wells. Mouse LEP in samples (or standards) combines with the coated antibody and HRP linked detection antibody special to Mouse LEP. Excess conjugate and unbound sample or standard are washed from the plate. The substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450±2 nm. The OD value is proportional to the concentration of Mouse LEP. The concentration of Mouse LEP in the samples is then determined by comparing the OD of the samples to the standard curve. | ||
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| (pg/mL) | OD | Corrected |
|---|---|---|
| 5000.00 | 2.400 | 2.340 |
| 2500.00 | 1.780 | 1.720 |
| 1250.00 | 1.032 | 0.972 |
| 625.00 | 0.523 | 0.463 |
| 312.50 | 0.271 | 0.211 |
| 156.25 | 0.146 | 0.086 |
| 78.13 | 0.096 | 0.036 |
| 0.00 | 0.060 |
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean(pg/mL) | 301.17 | 1056.44 | 2167.15 | 298.16 | 1098.70 | 2210.49 |
| Standard deviation | 16.11 | 33.91 | 134.36 | 18.01 | 33.51 | 134.84 |
| C V (%) | 5.35 | 3.21 | 6.20 | 6.04 | 3.05 | 6.10 |
The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 83-95 | 89 |
| EDTA plasma (n=8) | 95-107 | 101 |
| Cell culture media (n=8) | 82-94 | 88 |
Samples were spiked with high concentrations of target proteins and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 87-96 | 87-98 | 89-102 |
| Average (%) | 89 | 89 | 99 | |
| 1:4 | Range (%) | 82-95 | 87-98 | 82-97 |
| Average (%) | 91 | 91 | 85 | |
| 1:8 | Range (%) | 92-105 | 92-101 | 88-105 |
| Average (%) | 94 | 96 | 89 | |
| 1:16 | Range (%) | 89-97 | 82-98 | 83-96 |
| Average (%) | 94 | 83 | 90 |
