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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Mouse Lipocalin-2/NGAL Quantikine ELISA Kit
R&D Systems | Catalog # MLCN20
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (10 µL), Platelet-poor EDTA Plasma (10 µL), Platelet-poor Heparin Plasma (10 µL), Urine (10 µL)
Sensitivity
8.8 pg/mL
Assay Range
78.1-5000 pg/mL (Cell Culture Supernates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Urine)
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Product Summary for Mouse Lipocalin-2/NGAL Quantikine ELISA Kit
The Quantikine Mouse Lipocalin-2/NGAL Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse Lipocalin-2 in cell culture supernates, mouse serum, platelet-poor plasma, and urine. It contains NS0-expressed recombinant mouse Lipocalin-2 and antibodies raised against the recombinant factor. Natural mouse Lipocalin-2 showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards, indicating that this kit can be used to determine relative levels of natural mouse 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
Mouse
Specificity
Natural and recombinant mouse 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/Platelet-poor Plasma/Urine - Samples were evaluated for the presence of mouse
Lipocalin-2 in this assay.
Cell Culture Supernates:
Two mouse brains and kidneys were homogenized and seeded in RPMI 1640 supplemented with 10% fetal bovine serum, 5 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL of penicillin, and 100 μg/mL of streptomycin sulfate. The resulting supernates were either untreated or stimulated with 1 μg/mL of LPS for 3 days. Aliquots of the cell culture supernates were removed and assayed for levels of mouse Lipocalin-2.
| Sample Type | Mean (ng/mL) | Range (ng/mL) | Standard Deviation (ng/mL) |
| Serum (n=20) | 141 | 58.3-290 | 60.2 |
| Platelet-poor heparin plasma (n=20) | 76.0 | 40.7-143 | 30.1 |
| Platelet-poor EDTA plasma (n=20) | 54.1 | 32.5-74.7 | 12.3 |
| Urine (n=20) | 48.6 | 12.9-168 | 34.1 |
Cell Culture Supernates:
J774A.1 mouse reticulum cell sarcoma macrophage cells (1 x 106
cells/mL) were cultured in
RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL of penicillin,
and 100 μg/mL of streptomycin sulfate. Cells were either untreated or stimulated with
100 ng/mL of recombinant mouse IFN-gamma and 1 μg/mL of lipopolysaccharide (LPS) for 2 days.
Aliquots of the cell culture supernates were removed and assayed for levels of mouse
Lipocalin-2.
| Condition | Observed Values (ng/mL) |
| Unstimulated | 0.817 |
| Stimulated | 161 |
WEHI-3 myelomonocytic leukemia cells (2 x 105
cells/mL) were cultured in RPMI supplemented
with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL of penicillin, and 100 μg/mL of
streptomycin sulfate. Cells were either untreated or stimulated with 1 μg/mL of LPS for 4 days.
Aliquots of the cell culture supernates were removed and assayed for levels of mouse
Lipocalin-2.
| Condition | Observed Values (ng/mL) |
| Unstimulated | 83.7 |
| Stimulated | 178 |
Two mouse brains and kidneys were homogenized and seeded in RPMI 1640 supplemented with 10% fetal bovine serum, 5 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL of penicillin, and 100 μg/mL of streptomycin sulfate. The resulting supernates were either untreated or stimulated with 1 μg/mL of LPS for 3 days. Aliquots of the cell culture supernates were removed and assayed for levels of mouse Lipocalin-2.
| Condition | Observed Values (ng/mL) |
| Brain unstimulated | 0.334 |
| Brain stimulated | 178 |
| Condition | Observed Values (ng/mL) |
| Kidney Unstimulated | 1.28 |
| Kidney Stimulated | 4.69 |
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, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, 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) | 196 | 476 | 1534 | 211 | 477 | 1564 |
| Standard Deviation | 13.2 | 19.6 | 65.9 | 15.2 | 28.6 | 98.5 |
| CV% | 6.7 | 4.1 | 4.3 | 7.2 | 6.0 | 6.3 |
Recovery for Mouse Lipocalin-2/NGAL Quantikine ELISA Kit
The recovery of mouse Lipocalin-2 spiked to three levels throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Samples (n=4) | 93 | 82-101 |
Linearity
To assess the linearity of the assay, samples containing high concentrations of mouse Lipocalin-2 were diluted with Calibrator Diluent and then assayed.
Scientific Data Images for Mouse Lipocalin-2/NGAL Quantikine ELISA Kit
Mouse 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 Mouse 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 Mouse 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 Mouse Lipocalin-2/NGAL Quantikine ELISA Kit
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Sample Tested: SerumVerified Customer | Posted 12/05/2024Wide dynamic range. Easy to use kit.
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Sample Tested: SerumVerified Customer | Posted 12/11/2021
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Sample Tested: Kidney tissue and UrineVerified Customer | Posted 02/10/2021
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Sample Tested: SerumVerified Customer | Posted 01/07/2019Wild type (WT) or IL33-/- (Def) mice were given either vehicle (Veh) or cisplatin (Cis) to induce kidney injury. Serum NGAL was measured as a marker of kidney injury. Ravichandran et. al. AJP Renal 2017
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Sample Tested: SerumVerified Customer | Posted 07/19/2017Almost as good as human kit.
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Protocols
View specific protocols for Mouse Lipocalin-2/NGAL Quantikine ELISA Kit (MLCN20):
Refer to the product for complete assay procedure.
Bring all 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 50 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 room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 4 times.
- Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
50 µL Assay Diluent
50 µL Standard, Control, or Sample
100 µL Conjugate
100 µL Substrate Solution
100 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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