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Assay Type
Solid Phase Sandwich ELISA
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Format
96-well strip plate
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Assay Length
3.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Citrate Plasma (10 uL)
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Sensitivity
7.8 pg/mL
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Assay Range
15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Specificity
Natural and recombinant human Leptin
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
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Interference
Interference observed with 1 or more available related molecules.
Control Available
QC237, Quantikine Immunoassay Control Group 237 -
Product Summary
The Quantikine Human Leptin Immunoassay is a 3.5 hour solid phase ELISA designed to measure soluble human Leptin in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human Leptin and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant Leptin accurately. Results obtained measuring natural human Leptin showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human Leptin.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 64.5 | 146 | 621 | 65.7 | 146 | 581 |
| Standard Deviation | 2.14 | 4.32 | 20 | 3.56 | 6.17 | 20.6 |
| CV% | 3.3 | 3 | 3.2 | 5.4 | 4.2 | 3.5 |
Recovery
The recovery of Leptin spiked to three different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=5) | 98 | 94-102 |
| Citrate Plasma (n=5) | 95 | 87-105 |
| EDTA Plasma (n=5) | 99 | 85-112 |
| Heparin Plasma (n=5) | 90 | 81-100 |
| Serum (n=5) | 95 | 89-109 |
Linearity
To assess the linearity of the assay, cell culture media samples spiked with high concentrations of Leptin and natural serum/plasma samples were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: Leptin/OB
Leptin is a product of the mouse obese gene. Mice with mutations in the obese gene that block the synthesis of leptin are obese and diabetic, and have reduced activity, metabolism, and body temperature. Rat leptin shares approximately 96% and 82% sequence identity with the mouse and human protein, respectively. The expression of leptin mRNA is restricted to adipose tissue.
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Alternate Names
FLJ94114; LEP; leptin (murine obesity homolog); leptin (obesity homolog, mouse); Leptin; OB; Obese protein; obese, mouse, homolog of; Obesity factor; OBOBS;
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