Recombinant Human Lipocalin-2/NGAL Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Binding Activity
Format
Carrier-Free

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Recombinant Human 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. rhLipocalin-2 can bind >1.5 µM of Fe(DHBA)3 under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Lipocalin-2/NGAL protein
Gln21-Gly198, 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 under reducing conditions and visualized by silver stain.
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
25-27 kDa doublet, reducing conditions
Publications
Read Publications using
1757-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 under reducing conditions and visualized by silver stain.
Assay Procedure
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, pH 7.5 (TCN)
  • Recombinant Human Lipocalin‑2/NGAL (rhLipocalin-2) (Catalog # 1757-LC)
  • Iron III (Fe3+) (Sigma, Catalog # 16596)
  • 2,3,Dihydroxybenzoic Acid (DHBA) (Sigma, Catalog # 126209)
  • Ligand Buffer: 0.1 M Tris, pH 8.0
  • 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 containing 1 mM DHBA and Ligand Buffer.
    4. Incubate at room temperature for 10 minutes. A curve of the metal ligand complex of Fe(DHBA)3 is formed.
    5. After incubation, perform 5 fold dilutions to the curve using Assay Buffer.
    6. Dilute rhLipocalin-2 (MW: 21905 Da) to 4 µM in Assay Buffer.
    7. In the plate, load 50 µL of the diluted Fe(DHBA)3 complex curve and 50 µL of 4 µM rhLipocalin-2.
    8. Incubate 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 Curve: 0.001, 0.125, 0.25, 0.5, 1.0, 2.0, 4.0, 8.0, 16, and 32 µM.
  • rhLipocalin-2: 2 µM

Notes

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

Alternate Names for Recombinant Human 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

Members of Lipocalin family share a highly conserved fold with an eight-stranded antiparallel beta barrel, and act as a transporters, carrying small molecules to specific cells (1). Lipocalin-2, also known as Neutrophil Gelatinase-Associated Lipocalin (NGAL), was originally identified as a component of neutrophil granules (2). It is a 25 kDa protein existing in monomeric and homo- and heterodimeric forms, the latter as a dimer with human neutrophil gelatinases (MMP-9) (2). Its expression has been observed in most tissues normally exposed to microorganism, and its synthesis is induced in epithelial cells during inflammation (3). 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 sidropore bound to 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 by 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 epithelial to mesenchymal transition inducing activity of Ras, through restoration of E-cadherin expression, via effects on the Ras-MAPK signaling pathway (8).

  1. Flower, D.R. et al. (1994) FEBS Lett. 354:7.
  2. Kjeldsen, L. et al. (1993) J. Biol. Chem. 268:10426.
  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.

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Publications for Lipocalin-2/NGAL (1757-LC)(25)

We have publications tested in 5 confirmed species: Human, Mouse, Rat, Hamster, Salmonella.

We have publications tested in 7 applications: Bioassay, Cell Culture, ELISA (Standard), ELISA Developmet, Enzyme Assay, Stimulation, Western Blot.


Filter By Application
Bioassay
(18)
Cell Culture
(3)
ELISA (Standard)
(1)
ELISA Developmet
(1)
Enzyme Assay
(1)
Stimulation
(1)
Western Blot
(1)
All Applications
Filter By Species
Human
(20)
Mouse
(2)
Rat
(1)
Hamster
(1)
Salmonella
(1)
All Species
Showing Publications 1 - 10 of 25. Show All 25 Publications.
Publications using 1757-LC Applications Species
Kido, R;Hiroshima, Y;Kido, JI;Bando, M;Yoshida, K;Inagaki, Y;Yumoto, H; Lipocalin 2 inhibits the expressions of interleukin-8 and macrophage inflammatory protein-1? in human neutrophil-like cells Journal of oral biosciences 2025-01-30 [PMID: 39892784] (Bioassay, Human) Bioassay Human
Li, P;Chen, J;Wang, M;Wang, Q;Liu, X; High?fat diet?induced LCN2 exacerbates myocardial ischemia?reperfusion injury by enhancing platelet activation Molecular medicine reports 2024-11-01 [PMID: 39301623] (Bioassay, Mouse) Bioassay Mouse
W Santus, AP Rana, JR Devlin, KA Kiernan, CC Jacob, J Tjokrosurj, DM Underhill, J Behnsen Mycobiota and diet-derived fungal xenosiderophores promote Salmonella gastrointestinal colonization Nature Microbiology, 2022-11-21;7(12):2025-2038. 2022-11-21 [PMID: 36411353] (Bioassay, Salmonella) Bioassay Salmonella
U Gupta, S Ghosh, CT Wallace, P Shang, Y Xin, AP Nair, M Yazdankhah, A Strizhakov, MA Ross, H Liu, S Hose, NA Stepicheva, O Chowdhury, M Nemani, V Maddipatla, R Grebe, M Das, KL Lathrop, JA Sahel, JS Zigler, J Qian, A Ghosh, Y Sergeev, JT Handa, CM St Croix, D Sinha Increased LCN2 (lipocalin 2) in the RPE decreases autophagy and activates inflammasome-ferroptosis processes in a mouse model of dry AMD Autophagy, 2022-04-26;0(0):1-20. 2022-04-26 [PMID: 35473441] (Bioassay, Human) Bioassay Human
L Cook, M Sengelmann, B Winkler, C Nagl, S Koch, U Schlomann, EP Slater, MA Miller, EP von Strand, B Dörsam, C Preu beta er, JW Bartsch ADAM8-Dependent Extracellular Signaling in the Tumor Microenvironment Involves Regulated Release of Lipocalin 2 and MMP-9 International Journal of Molecular Sciences, 2022-02-10;23(4):. 2022-02-10 [PMID: 35216088] (Stimulation, Human) Stimulation Human
SK Schröder, M Pinoé-Schm, R Weiskirche Lipocalin-2 (LCN2) Deficiency Leads to Cellular Changes in Highly Metastatic Human Prostate Cancer Cell Line PC-3 Cells, 2022-01-13;11(2):. 2022-01-13 [PMID: 35053376] (Bioassay, Human) Bioassay Human
W Zhang, R Pan, M Lu, Q Zhang, Z Lin, Y Qin, Z Wang, S Gong, H Lin, S Chong, L Lu, W Liao, X Lu Epigenetic induction of lipocalin 2 expression drives acquired resistance to 5-fluorouracil in colorectal cancer through integrin beta3/SRC pathway Oncogene, 2021-09-29;0(0):. 2021-09-29 [PMID: 34588619] (Bioassay, Human) Bioassay Human
X Wen, B Su, M Gao, J Chen, D Zhou, H You, N Li, S Chang, X Cheng, C Qian, J Gao, P Yang, S Qu, L Bu Obesity-associated up-regulation of lipocalin 2 protects gastric mucosa cells from apoptotic cell death by reducing endoplasmic reticulum stress Cell Death & Disease, 2021-02-26;12(2):221. 2021-02-26 [PMID: 33637683] (Bioassay, Human) Bioassay Human
A Tyagi, S Sharma, K Wu, SY Wu, F Xing, Y Liu, D Zhao, RP Deshpande, RB D'Agostino, K Watabe Nicotine promotes breast cancer metastasis by stimulating N2 neutrophils and generating pre-metastatic niche in lung Nature Communications, 2021-01-20;12(1):474. 2021-01-20 [PMID: 33473115] (Bioassay, Human) Bioassay Human
Z Hui, Z Jiang, D Qiao, Z Bo, K Qiyuan, B Shaohua, Y Wenbing, L Wei, L Cheng, L Shuangning, L Zhengyi, L Yingyi Increased expression of LCN2 formed a positive feedback loop with activation of the ERK pathway in human kidney cells during kidney stone formation Scientific Reports, 2020-12-04;10(1):21287. 2020-12-04 [PMID: 33277533] (Cell Culture, Human) Cell Culture Human
Show All 25 Publications.

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Bioinformatics

Gene Symbol LCN2
Uniprot