Mouse Lipocalin-2 was cloned from mouse kidney cells (1). Its very high level of expression at the post-stratum uterus gave it the name uterocalin (2). Lipocalin-2 has been implicated in a variety of processes including cell differentiation, tumorigenesis, and apoptosis (3‑5). Studies indicate that Lipocalin-2 binds a bacterial catecholate siderophore that is bound to a ferric ion, such as enterobactin, with a subnanomolar dissociation constant (KD = 0.41 nM) (6). The bound ferric enterobactin complex breaks down slowly in a month into dihydroxybenzoyl serine and dihydroxybenzoic acid (DHBA). It also binds to a ferric DHBA complex with much less KD values (7.9 nM) (6). Secretion of Lipocalin-2 in immune cells increases in response to stimulation of Toll-like receptor as an acute phase response to infection. As a result, it acts as a potent bacteriostatic reagent by sequestering iron (7). Moreover, Lipocalin-2 can alter the invasive and metastatic behavior of Ras-transformed breast cancer cells in vitro and in vivo by reversing the epithelial to mesenchymal transition inducing activity of Ras, through restoration of E-cadherin expression, via effects on the Ras-MAPK signaling pathway (8).
Recombinant Mouse Lipocalin-2/NGAL Protein, CF
R&D Systems | Catalog # 1857-LC
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Lipocalin-2/NGAL Protein (1857-LC)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
NS0
Accession Number
Applications
Binding Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived mouse Lipocalin-2/NGAL protein
Gln21-Asn200 with a C-terminal 10 His tag
Gln21-Asn200 with a C-terminal 10 His tag
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
No results obtained: Gln21 predicted
Predicted Molecular Mass
22 kDa
SDS-PAGE
26-27 kDa doublet, reducing conditions
Activity
Measured by its ability to bind Iron(III) dihydroxybenzoic acid [Fe(DHBA)3]. The binding of Fe(DHBA)3 results in the quenching of Trp fluorescence in Lipocalin-2.
>1.0 µM of Fe(DHBA)3 can be bound under the described conditions.
>1.0 µM of Fe(DHBA)3 can be bound under the described conditions.
Reviewed Applications
Read 1 review rated 5 using 1857-LC in the following applications:
Formulation, Preparation, and Storage
1857-LC
| Formulation | Supplied as a 0.2 μm filtered solution in MES and NaCl. |
| Shipping | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: Lipocalin-2/NGAL
References
- Hraba-Renevey, s. et al. (1989) Oncogene. 4:601.
- Liu, Q. et al. (1993) Mol Reprod Dev. 46:507.
- Kjeldsen L, et al. (2000) Biochim Biophys Acta. 1482:272.
- Devireddy, L.R. et al. (2001) Science 293:829.
- Yang, M.B. et al. (2002) Mol. Cell. 10:1045.
- Goetz, D.H. et al. (2002) Mol. Cell 10:1033.
- Flo, T.H. et al. (2004) Nature 432:917.
- Hanai, J. et al. (2005) J. Biol. Chem. 280:13641.
Long Name
Neutrophil Gelatinase-associated Lipocalin
Alternate Names
24p3, LCN2, Lipocalin2, MSFI, NGAL, Oncogene 24p3, p25, Siderocalin, Uterocalin
Gene Symbol
LCN2
UniProt
Additional Lipocalin-2/NGAL Products
Product Documents for Recombinant Mouse Lipocalin-2/NGAL Protein, CF
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Mouse Lipocalin-2/NGAL Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Mouse Lipocalin-2/NGAL Protein, CF
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Protocols
View specific protocols for Recombinant Mouse Lipocalin-2/NGAL Protein, CF (1857-LC):
Materials
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, pH 7.5 (TCN)
- Ligand Buffer: 0.1 M Tris, pH 8.0
- Recombinant Mouse Lipocalin‑2/NGAL (rmLipocalin‑2) (Catalog # 1857-LC)
- Iron III (Fe3+) (Sigma, Catalog # 16596)
- 2,3,Dihydroxybenzoic Acid (DHBA) (Sigma, Catalog # 126209)
- F16 Black Maxisorp Plate (Nunc, Catalog #: 475515)
- Fluorescent Plate Reader (Model: Spectramax Gemini EM by Molecular Devices) or equivalent
- Prepare a curve of Fe3+ in deionized water with the following serial dilutions: 640, 320, 160, 80, 40, 20, 10, 5, and 2.5 µM.
- Prepare 1 mM DHBA in Ligand Buffer from powder stock.
- Combine equal volumes of the Fe3+ curve with 1 mM DHBA. Include a control of 1 mM DHBA and Ligand Buffer.
- Incubate at room temperature for 10 minutes.
- After incubation, perform 5 fold dilutions to the curve with Assay Buffer.
- Dilute rmLipocalin-2 (MW: 21905 Da) to 4 µM in Assay Buffer.
- In a plate, load 50 µL of the diluted Fe(DHBA)3 complex curve and 50 µL of 4 µM rmLipocalin-2.
- Incubate reaction at room temperature for 30 minutes.
- Read at excitation and emission wavelengths of 280 nM and 340 nM, respectively in endpoint mode.
- Plot a 4-parameter curve of Fe(DHBA)3 concentration (x-axis) versus RFUs (y-axis), and calculate a BC50 from the curve.
- Fe(DHBA)3 complex: 0.125, 0.25, 0.5, 1.0, 2.0, 4.0, 8.0, 16, and 32 µM.
- rmLipocalin-2: 2 µM
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