Leptin, also known as OB, is a polypeptide hormone that regulates adipose tissue mass and energy balance. In mice (ob/ob) and humans, inactivating mutations of the OB gene can cause obesity. Leptin signals through the transmembrane Leptin R in the hypothalamus but can also bind shorter and soluble forms of Leptin R which do not signal. Adipocytes increase Leptin expression as cell size increases, resulting in depressed appetite and increased energy expenditure. Low Leptin levels prompt increased appetite and decreased energy expenditure. Leptin can also block the onset of puberty or menses during starvation, extreme exercise-induced weight loss, anorexia, or cancer-induced cachexia.
Human Leptin Quantikine QuicKit ELISA
R&D Systems | Catalog # QK398
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Key Product Details
Assay Length
80 minutes
Sample Type & Volume Required Per Well
Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL)
Sensitivity
1.03 pg/mL
Assay Range
15.6-1000 pg/mL (Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Human Leptin Quantikine QuicKit ELISA
The Quantikine® QuicKit™ Human Leptin Immunoassay is a one step, 80-minute solid phase ELISA designed to measure human Leptin levels in cell culture supernates, serum, and plasma. It contains E.
coli-expressed recombinant human Leptin and antibodies raised against the recombinant protein. Results obtained using natural human Leptin showed linear curves that were parallel to the standard curves obtained using the QuicKit™ standards. These results indicate that this kit can be used to determine relative mass values for natural human Leptin.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
Natural and recombinant human Leptin.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Precision
Intra-Assay Precision (Precision within an assay) Two samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Two samples of known concentration were tested in ten separate assays to assess inter-assay precision. Assays were performed by at least three technicians.
EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||
|---|---|---|---|---|
| Sample | 1 | 2 | 1 | 2 |
| n | 20 | 20 | 10 | 10 |
| Mean (pg/mL) | 134 | 709 | 130 | 709 |
| Standard Deviation | 2.84 | 15.8 | 11.9 | 19.7 |
| CV% | 2.1 | 2.2 | 9.2 | 2.8 |
Recovery for Human Leptin Quantikine QuicKit ELISA
The recovery of human Leptin spiked to three levels in samples throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 89 | 80-106 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Leptin were diluted with calibrator diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Scientific Data Images for Human Leptin Quantikine QuicKit ELISA
Human Leptin ELISA Standard Curve
Human Leptin QuicKit Spiked Recovery Competitor Comparison
Leptin is spiked at three known concentrations throughout the range of the assay and run to measure response of the spiked sample matrix. Serum recovery is 74% compared to 70% for the top competitor. EDTA plasma recovery is 84% compared to 69% for the top competitor. Heparin plasma recovery is 88% compared to 77% for the top competitor. In spike and recovery experiments, natural samples are spiked with the recombinant target analyte of interest to identify interference caused by sample matrices.
Human Leptin QuicKit Linearity Competitor Comparison
Samples containing and/or spiked with high concentrations of Leptin in various matrices and diluted with appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay. The linearity is between 99%-106% compared to 71%-122% for the top competitor.Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Leptin/OB
Additional Leptin/OB Products
Product Documents for Human Leptin Quantikine QuicKit ELISA
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Leptin Quantikine QuicKit ELISA
For research use only
Citations for Human Leptin Quantikine QuicKit ELISA
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Protocols
View specific protocols for Human Leptin Quantikine QuicKit ELISA (QK398):
These assays utilize an anti-tag coated microplate. Standards, samples, and controls are added to plate, followed by subsequent addition of an antibody cocktail. Following a 1 hour incubation, plates are washed and substrate is added and incubated for 20 minutes. Stop solution is then added and plates are read using a standard microplate reader.
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