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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
4.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (20 uL), EDTA Plasma (20 uL)
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Sensitivity
22 pg/mL
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Assay Range
62.5 - 4,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
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Specificity
Natural and recombinant mouse and rat Leptin. This assay cross-reacts 0.24% with Human Leptin.
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity observed with 1 or more species tested.
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Interference
Interference observed with 1 or more available related molecules.
Product Summary
The Quantikine Mouse Leptin Immunoassay is a 4.5 hour solid phase ELISA designed to measure soluble mouse leptin in cell culture supernates, serum*, and plasma. It contains recombinant mouse leptin expressed in E. coli and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse leptin accurately. Results obtained measuring natural mouse leptin showed dose-response curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse leptin.
*Pregnant mouse serum from the second trimester of pregnancy through post-partum day 1 require acid/urea activation of mouse 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 | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 182 | 693 | 1803 | 187 | 654 | 1736 |
| Standard Deviation | 7.9 | 22.6 | 67.9 | 14.2 | 30.5 | 87.4 |
| CV% | 4.3 | 3.3 | 3.8 | 7.6 | 4.7 | 5 |
Recovery
The recovery of mouse/rat Leptin spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Mouse Cell Culture Supernates (n=5) | 101 | 85-120 |
| Mouse EDTA Plasma (n=4) | 92 | 78-105 |
| Mouse Heparin Plasma (n=4) | 95 | 81-105 |
| Mouse Serum (n=18) | 108 | 81-122 |
| Rat Cell Culture Supernates (n=4) | 114 | 103-125 |
| Rat EDTA Plasma (n=7) | 96 | 83-114 |
| Rat Serum (n=10) | 92 | 81-115 |
Linearity
To assess the linearity of the assay, four or more samples containing and/or spiked with high concentrations of mouse/rat Leptin in each matrix were diluted with Calibrator Diluent and then assayed.
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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