Recombinant Mouse Lipocalin-2/NGAL Protein, CF

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Summary
Reactivity MuSpecies Glossary
Applications Bind
Format
Carrier-Free

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Recombinant Mouse Lipocalin-2/NGAL Protein, CF Summary

Details of Functionality
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.
Source
Mouse myeloma cell line, NS0-derived
Gln21-Asn200 with a C-terminal 10 His tag
Accession #
N-terminal Sequence
No results obtained: Gln21 predicted
Protein/Peptide Type
Recombinant Enzymes
Gene
Lcn2
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
22 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
26-27 kDa doublet, reducing conditions
Publications
Read Publications using
1857-LC in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in MES and NaCl.
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • 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
  1. 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.
  2. Prepare 1 mM DHBA in Ligand Buffer from powder stock.
  3. Combine equal volumes of the Fe3+ curve with 1 mM DHBA. Include a control of 1 mM DHBA and Ligand Buffer.
  4. Incubate at room temperature for 10 minutes.
  5. After incubation, perform 5 fold dilutions to the curve with Assay Buffer.
  6. Dilute rmLipocalin-2 (MW: 21905 Da) to 4 µM in Assay Buffer.
  7. In a plate, load 50 µL of the diluted Fe(DHBA)3 complex curve and 50 µL of 4 µM rmLipocalin-2.
  8. Incubate reaction at room temperature for 30 minutes.
  9. Read at excitation and emission wavelengths of 280 nM and 340 nM, respectively in endpoint mode.
  10. Plot a 4-parameter curve of Fe(DHBA)3 concentration (x-axis) versus RFUs (y-axis), and calculate a BC50 from the curve.
Per Well:
  • 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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse Lipocalin-2/NGAL Protein, CF

  • 24p3
  • 25 kDa alpha-2-microglobulin-related subunit of MMP-9
  • HNL
  • LCN2
  • lipocalin 2 (oncogene 24p3)
  • lipocalin 2
  • Lipocalin2
  • Lipocalin-2
  • migration-stimulating factor inhibitor
  • MSFI
  • neutrophil gelatinase-associated lipocalin
  • NGAL
  • NGALlipocalin-2
  • Oncogene 24p3
  • p25
  • Siderocalin

Background

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).

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Publications for (9)

We have publications tested in 1 confirmed species: Mouse.

We have publications tested in 2 applications: Bioassay, In Vivo.


Filter By Application
Bioassay
(6)
In Vivo
(3)
All Applications
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Mouse
(9)
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Showing Publications 1 - 9 of 9.
Publications using 1857-LC Applications Species
AC Ferreira, T Santos, B Sampaio-Ma, A Novais, SD Mesquita, P Ludovico, L Bernardino, M Correia-Ne, N Sousa, JA Palha, JC Sousa, F Marques Lipocalin-2 regulates adult neurogenesis and contextual discriminative behaviours Mol. Psychiatry, 2017;0(0):. 2017 [PMID: 28485407] (Bioassay, Mouse) Bioassay Mouse
The adipokine lipocalin-2 in the context of the osteoarthritic osteochondral junction Sci Rep, 2016;6(0):29243. 2016 [PMID: 27385438] (Bioassay, Mouse) Bioassay Mouse
Rucci N, Capulli M, Piperni S, Cappariello A, Lau P, Frings-Meuthen P, Heer M, Teti A Lipocalin 2: a new mechanoresponding gene regulating bone homeostasis. J Bone Miner Res, 2015;30(2):357-68. 2015 [PMID: 25112732] (Bioassay, Mouse) Bioassay Mouse
Maung R, Hoefer M, Sanchez A, Sejbuk N, Medders K, Desai M, Catalan I, Dowling C, de Rozieres C, Garden G, Russo R, Roberts A, Williams R, Kaul M CCR5 knockout prevents neuronal injury and behavioral impairment induced in a transgenic mouse model by a CXCR4-using HIV-1 glycoprotein 120. J Immunol, 2014;193(4):1895-910. 2014 [PMID: 25031461] (Bioassay, Mouse) Bioassay Mouse
Miao Q, Ku A, Nishino Y, Howard J, Rao A, Shaver T, Garcia G, Le D, Karlin K, Westbrook T, Poli V, Nguyen H Tcf3 promotes cell migration and wound repair through regulation of lipocalin 2. Nat Commun, 2014;5(0):4088. 2014 [PMID: 24909826] (In Vivo, Mouse) In Vivo Mouse
Jeon, Sangmin, Jha, Mithiles, Ock, Jiyeon, Seo, Jungwan, Jin, Myungwon, Cho, Heejung, Lee, Won-Ha, Suk, Kyoungho Role of lipocalin-2-chemokine axis in the development of neuropathic pain following peripheral nerve injury. J Biol Chem, 2013;288(33):24116-27. 2013 [PMID: 23836894] (In Vivo, Mouse) In Vivo Mouse
Shashidharamurthy R, Machiah D, Aitken J, Putty K, Srinivasan G, Chassaing B, Parkos C, Selvaraj P, Vijay-Kumar M Differential role of lipocalin 2 during immune complex-mediated acute and chronic inflammation in mice. Arthritis Rheum, 2013;65(4):1064-73. 2013 [PMID: 23280250] (In Vivo, Mouse) In Vivo Mouse
Nairz M, Theurl I, Schroll A, Theurl M, Fritsche G, Lindner E, Seifert M, Crouch ML, Hantke K, Akira S, Fang FC, Weiss G Absence of functional Hfe protects mice from invasive Salmonella enterica serovar Typhimurium infection via induction of lipocalin-2. Blood, 2009;114(17):3642-51. 2009 [PMID: 19700664] (Bioassay, Mouse) Bioassay Mouse
Zhang J, Wu Y, Zhang Y, LeRoith D, Bernlohr DA, Chen X The role of lipocalin 2 in the regulation of inflammation in adipocytes and macrophages. Mol. Endocrinol., 2008;22(6):1416-26. 2008 [PMID: 18292240] (Bioassay, Mouse) Bioassay Mouse

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Bioinformatics

Gene Symbol Lcn2
Entrez
Uniprot