Recombinant Mouse Lipocalin-2/NGAL Protein, CF

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Product Details

Summary
Reactivity MuSpecies Glossary
Applications Binding Activity
Format
Carrier-Free

Order Details

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 mouse Lipocalin-2/NGAL protein
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

Dilutions
  • Binding Activity
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
  • Uterocalin

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).

  1. Hraba-Renevey, s. et al. (1989) Oncogene. 4:601. 
  2. Liu, Q. et al. (1993) Mol Reprod Dev. 46:507. 
  3. Kjeldsen L, et al. (2000) Biochim Biophys Acta. 1482:272.
  4. Devireddy, L.R. et al. (2001) Science 293:829.
  5. Yang, M.B. et al. (2002) Mol. Cell. 10:1045.
  6. Goetz, D.H. et al. (2002) Mol. Cell 10:1033.
  7. Flo, T.H. et al. (2004) Nature 432:917.
  8. Hanai, J. et al. (2005) J. Biol. Chem. 280:13641.
  9.  

Publications for Lipocalin-2/NGAL (1857-LC)(23)

We have publications tested in 3 confirmed species: Human, Mouse, Rat.

We have publications tested in 3 applications: Bioassay, Cell Culture, In Vivo.


Filter By Application
Bioassay
(13)
Cell Culture
(1)
In Vivo
(9)
All Applications
Filter By Species
Human
(1)
Mouse
(20)
Rat
(1)
All Species
Showing Publications 1 - 10 of 23. Show All 23 Publications.
Publications using 1857-LC Applications Species
Su, H;Guo, H;Qiu, X;Lin, TY;Qin, C;Celio, G;Yong, P;Senders, M;Han, X;Bernlohr, DA;Chen, X; Lipocalin 2 regulates mitochondrial phospholipidome remodeling, dynamics, and function in brown adipose tissue in male mice Nature communications 2023-10-23 [PMID: 37872178] (Bioassay, Mouse) Bioassay Mouse
Liu, Y;Smith, MR;Wang, Y;D'Agostino, R;Ruiz, J;Lycan, T;Kucera, GL;Miller, LD;Li, W;Chan, MD;Farris, M;Su, J;Song, Q;Zhao, D;Chandrasekaran, A;Xing, F; c-Met Mediated Cytokine Network Promotes Brain Metastasis of Breast Cancer by Remodeling Neutrophil Activities Cancers 2023-05-05 [PMID: 37174093]
JR Sheldon, LE Himmel, DE Kunkle, AJ Monteith, KN Maloney, EP Skaar Lipocalin-2 is an essential component of the innate immune response to Acinetobacter baumannii infection PloS Pathogens, 2022-09-02;18(9):e1010809. 2022-09-02 [PMID: 36054235] (Bioassay, Mouse) Bioassay Mouse
X Xiang, X Tang, Y Yu, S Xie, L Liu, M Chen, R Zhang, X Kang, Y Zheng, G Yang, S Gan, S Zhu Role of lipocalin-2 in surgery-induced cognitive decline in mice: a signal from neuron to microglia Journal of Neuroinflammation, 2022-04-12;19(1):92. 2022-04-12 [PMID: 35413913] (Bioassay, In Vivo, Mouse) Bioassay, In Vivo Mouse
C Huisman, MA Norgard, PR Levasseur, SM Krasnow, MGP van der Wi, B Olson, DL Marks Critical changes in hypothalamic gene networks in response to pancreatic cancer as found by single-cell RNA sequencing Molecular Metabolism, 2022-01-11;0(0):101441. 2022-01-11 [PMID: 35031523] (In Vivo, Mouse) In Vivo Mouse
X Wang, X Li, X Zuo, Z Liang, T Ding, K Li, Y Ma, P Li, Z Zhu, C Ju, Z Zhang, Z Song, H Quan, J Zhang, X Hu, Z Wang Photobiomodulation inhibits the activation of neurotoxic microglia and astrocytes by inhibiting Lcn2/JAK2-STAT3 crosstalk after spinal cord injury in male rats Journal of Neuroinflammation, 2021-11-05;18(1):256. 2021-11-05 [PMID: 34740378] (Bioassay, Rat) Bioassay Rat
YH Chen, SY Xie, CW Chen, DY Lu Electroacupuncture improves repeated social defeat stress-elicited social avoidance and anxiety-like behaviors by reducing Lipocalin-2 in the hippocampus Molecular Brain, 2021-09-26;14(1):150. 2021-09-26 [PMID: 34565419] (In Vivo, Mouse) In Vivo Mouse
B Olson, X Zhu, MA Norgard, P Diba, PR Levasseur, AC Buenafe, C Huisman, KG Burfeind, KA Michaelis, G Kong, T Braun, DL Marks Chronic cerebral lipocalin 2 exposure elicits hippocampal neuronal dysfunction and cognitive impairment Brain, Behavior, and Immunity, 2021-07-08;0(0):. 2021-07-08 [PMID: 34245812] (In Vivo, Mouse) In Vivo Mouse
B Olson, X Zhu, MA Norgard, PR Levasseur, JT Butler, A Buenafe, KG Burfeind, KA Michaelis, KR Pelz, H Mendez, J Edwards, SM Krasnow, AJ Grossberg, DL Marks Lipocalin 2 mediates appetite suppression during pancreatic cancer cachexia Nature Communications, 2021-04-06;12(1):2057. 2021-04-06 [PMID: 33824339] (Bioassay, Mouse) Bioassay Mouse
J Yan, SP Pandey, BJ Barnes, JR Turner, C Abraham T Cell-Intrinsic IRF5 Regulates T Cell Signaling, Migration, and Differentiation and Promotes Intestinal Inflammation Cell Rep, 2020-06-30;31(13):107820. 2020-06-30 [PMID: 32610123] (Bioassay, Mouse) Bioassay Mouse
Show All 23 Publications.

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Bioinformatics

Gene Symbol Lcn2
Uniprot