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 DuoSet ELISA
R&D Systems | Catalog # DY1857
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Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
7.81-500 pg/mL
Sample Type
Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Reactivity
Mouse
Mouse Lipocalin-2/NGAL DuoSet ELISA Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
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Product Summary for Mouse Lipocalin-2/NGAL DuoSet ELISA
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse Lipocalin-2. The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Specifications
Assay Format
96-well strip plate (sold separately)
Sample Volume Required
100 µL
Detection Method
Colorimetric ELISA - 450nm (TMB)
Conjugate
Biotin
Specificity
Label
HRP
Scientific Data Images for Mouse Lipocalin-2/NGAL DuoSet ELISA
Mouse Lipocalin-2 / NGAL ELISA Standard Curve
Kit Contents for Mouse Lipocalin-2/NGAL DuoSet ELISA
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008C) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or equivalent
Reagent Diluent*
Blocking Buffer*
Substrate Solution: ELISA TMB Substrate (Catalog # DY999B or DY999B-250)
Stop Solution: Methanesulfonic acid (Catalog # DY994B or DY994B-250)
Microplates: (Catalog # DY990), or equivalent
Plate Sealers: (Catalog # DY992), or equivalent
*For the recommended Reagent Diluent and Blocking Buffer for a specific DuoSet ELISA Development Kit, refer to the product datasheet.
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 DuoSet ELISA
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 DuoSet ELISA
For research use only
Related Research Areas
Citations for Mouse Lipocalin-2/NGAL DuoSet ELISA
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Customer Reviews for Mouse Lipocalin-2/NGAL DuoSet ELISA (10)
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Sample Tested: Mouse fecal and mouse fecal sampleVerified Customer | Posted 01/23/2025Standard curve for Mouse Lipocalin-2/NGAL DuoSet ELISA
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Sample Tested: SerumVerified Customer | Posted 02/08/2023Great kit as usual
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Sample Tested: UrineVerified Customer | Posted 11/17/2022Great kit as usual. Always good results. We use 200x dilution for control samples and 200.000x dilution for AKI samples.
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Sample Tested: Serum and UrineVerified Customer | Posted 09/14/2020Fantastic Duoset kit. Verified with Serum and Urine. For urine I ran 1:200 to 1:100 and for serum I ran 1:400 to 1:600 but once again it all depends on your individual samples. Acceptable CVs on all 5 plates that I ran.
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Sample Tested: Frozen fecesVerified Customer | Posted 08/05/2020Dilution of the standard was unclear Everything else was fine
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Sample Tested: Mouse fecesVerified Customer | Posted 03/06/2020
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Sample Tested: frozen fecal sampleVerified Customer | Posted 01/14/2019
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Sample Tested: mouse serum, Cell Culture Supernates, murine serum and murine tissue lysatesVerified Customer | Posted 12/07/2017White Adipose Tissue (WAT) and Brown Adipose Tissue (BAT) lysates from wild-type mice. 500 ng total protein, 3-4 mice.
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Sample Tested: Colon tissueVerified Customer | Posted 04/21/2017
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Sample Tested: Serum and fecal extractVerified Customer | Posted 01/15/2016None: Every detail available with kit and very easy to use.
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Protocols
View specific protocols for Mouse Lipocalin-2/NGAL DuoSet ELISA (DY1857):
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
- ELISA Troubleshooting Guide
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- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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