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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 (50 uL), Heparin Plasma (50 uL), Saliva (50 uL), Urine (50 uL)
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Sensitivity
0.04 ng/mL
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Assay Range
0.2 - 10 ng/mL (Cell Culture Supernates, Serum, Heparin Plasma, Saliva, Urine)
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Specificity
Natural and recombinant human Lipocalin-2
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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
No significant interference observed with available related molecules.
Control Available
QC115, Quantikine Immunoassay Control Set 736 for Human Lipocalin-2 -
Product Summary
The Quantikine Human Lipocalin-2 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Lipocalin-2 in cell culture supernates, serum, plasma, urine, and saliva. It contains NS0-expressed recombinant human Lipocalin-2 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Lipocalin-2 showed linear 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 naturally occurring human Lipocalin-2.
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, Heparin Plasma, Saliva, Urine | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 1.14 | 3.45 | 7.54 | 1.05 | 3.37 | 6.92 |
| Standard Deviation | 0.041 | 0.107 | 0.334 | 0.083 | 0.204 | 0.387 |
| CV% | 3.6 | 3.1 | 4.4 | 7.9 | 6.1 | 5.6 |
Recovery
The recovery of Lipocalin-2 spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
| Cell Culture Media (n=4) | 93 | 88-101 |
| Urine (n=4) | 90 | 84-97 |
Linearity
To assess the linearity of the assay, samples containing high concentrations of Lipocalin-2 were serially diluted with the Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Background: Lipocalin-2/NGAL
The Lipocalin family comprises a diverse group of mostly secreted soluble proteins that bind hydrophobic ligands and act as transporters, carrying small molecules to specific cells. Lipocalins are related by possessing an 8-stranded beta-barrel structure. Lipocalin-1, also named tear lipocalin (TL), von Ebners gland protein (VEG) and tear pre-albumin, binds a large number of hydrophobic molecules and exhibits cysteine proteinase inhibitor and endonuclear activities. Lipocalin-2, also known as neutrophil gelatinase-associated lipocalin (NGAL), is a component of granules in neutrophils from tissues that are normally exposed to microorganisms and is upregulated during inflammation. Lipocalin-2 can form homodimers and can heterodimerize with the neutrophil gelatinase MMP-9.
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