Lipocalin-2/NGAL (Neutrophil Gelatinase-associated Lipocalin) is a secreted carrier protein that acts in the innate immune response, differentiation, tumorigenesis, and cell survival. It is induced in macrophages, glial cells, and epithelial cells during inflammation, anemia, and hypoxia. Lipocalin-2 binds catecholate siderophores which are secreted by bacteria and are required for bacterial iron uptake. Lipocalin-2 functions as a bacteriostatic agent in the innate immune response by limiting the bacterial iron supply. Iron uptake by Lipocalin-2 into mammalian cells is important for the regulation of iron-sensitive gene transcription. In the kidney, Lipocalin-2-mediated iron trafficking is required for protection from renal injury.
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Human Lipocalin-2/NGAL Quantikine ELISA Kit
R&D Systems | Catalog # DLCN20
Quantikine Quickit version available - results in 90 minutes (QK1757).
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), Heparin Plasma (50 µL), Saliva (50 µL), Urine (50 µL)
Sensitivity
0.04 ng/mL
Assay Range
0.156-10 ng/mL (Cell Culture Supernates, Serum, Heparin Plasma, Saliva, Urine)
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Product Summary for Human Lipocalin-2/NGAL Quantikine ELISA Kit
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.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
Natural and recombinant human Lipocalin-2
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.
Sample Values
Serum/Plasma/Saliva/Urine - Samples from apparently healthy volunteers were evaluated
for the presence of human Lipocalin-2 in this assay. No medical histories were available for the
donors used in this study.
A431human epithelial carcinoma cells were grown to confluency in DMEM (high glucose) with 10% fetal bovine serum. An aliquot of the cell culture supernate was removed, assayed for human Lipocalin-2, and measured 130 ng/mL.
| Sample Type | Mean (ng/mL) | Range (ng/mL) | Standard Deviation (ng/mL) |
| Serum (n=35) | 119 | 42-177 | 35.6 |
| Heparin plasma (n=35) | 94 | 36-149 | 28.7 |
| Saliva (n=9) | 320 | 96-881 | 238 |
| urine (n=19) | 9.94 | 0.40-72 | 17.1 |
Cell Culture Supernates:
Human peripheral blood cells (1 x 106
cells/mL) were cultured in DMEM supplemented
with 5% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin,
and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with
10 μg/mL PHA. Aliquots of the cell culture supernates were removed and assayed for levels of
human Lipocalin-2.
| Condition | Day 1 (ng/mL) | Day 5 (ng/mL) |
| Unstimulated | 7.73 | 7.73 |
| Stimulated | 14.3 | 18.5 |
A431human epithelial carcinoma cells were grown to confluency in DMEM (high glucose) with 10% fetal bovine serum. An aliquot of the cell culture supernate was removed, assayed for human Lipocalin-2, and measured 130 ng/mL.
Neutrophils were cultured at 1 x 107
cells/mL in Hank's media and stimulated with 50 ng/mL
PMA for 30 minutes. An aliquot of the cell culture supernate was removed, assayed for human
Lipocalin-2, and measured 699 ng/mL.
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, Heparin Plasma, Saliva, Serum, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 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 for Human Lipocalin-2/NGAL Quantikine ELISA Kit
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.
Scientific Data Images for Human Lipocalin-2/NGAL Quantikine ELISA Kit
Human Lipocalin-2/NGAL ELISA Standard Curve
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: Lipocalin-2/NGAL
Long Name
Neutrophil Gelatinase-associated Lipocalin
Alternate Names
24p3, LCN2, Lipocalin2, MSFI, NGAL, Oncogene 24p3, p25, Siderocalin, Uterocalin
Gene Symbol
LCN2
Additional Lipocalin-2/NGAL Products
Product Documents for Human Lipocalin-2/NGAL Quantikine ELISA Kit
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 Lipocalin-2/NGAL Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Human Lipocalin-2/NGAL Quantikine ELISA Kit
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Sample Tested: Human retinaVerified Customer | Posted 02/24/2023The quantification of human lipocalin-2/NGAL was used in retina obtained from a clinical study quantifying diabetic retinopathy. In this model, the disease samples from patients displaying retinopathy were used as those having the pro-inflammatory effect. In the vitreous compartment, lysis of this tissue was used to quantify human lipocalin-2.
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Sample Tested: UrineVerified Customer | Posted 10/24/2020Porcine urine samples
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Sample Tested: UrineVerified Customer | Posted 04/24/2018Most urine samples fell within the curve at a 1 to 2 dilution. The higher NGAL samples required dilution at 1 to 10
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Sample Tested: SerumVerified Customer | Posted 07/18/2017Performed ideally and is probably the best kit for analyzing lipocalin2 out there! Although results should be analyzed at once after completion as saving the ELISA plate over night creates dark spots inside of the wells (see picture/data). Don´t forget to dilute serum!
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Protocols
View specific protocols for Human Lipocalin-2/NGAL Quantikine ELISA Kit (DLCN20):
Refer to the product for complete assay procedure.
The conjugate must remain at 2-8 °C during use. Bring all other reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at 2-8 °C for 2 hours.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 2 hours.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
100 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL cold Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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