Recombinant Mouse Lipocalin-2/NGAL Protein, CF

Catalog # Availability Size / Price Qty
1857-LC-050
R&D Systems Recombinant Proteins and Enzymes
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Product Details
Citations (19)
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Recombinant Mouse Lipocalin-2/NGAL Protein, CF Summary

Product Specifications

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.
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.
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
Analysis
No results obtained: Gln21 predicted
Predicted Molecular Mass
22 kDa
SDS-PAGE
26-27 kDa doublet, reducing conditions

Product Datasheets

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1857-LC

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

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
  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
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Background: Lipocalin-2/NGAL

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).

References
  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.  
Long Name
Neutrophil Gelatinase-associated Lipocalin
Entrez Gene IDs
3934 (Human); 16819 (Mouse); 170496 (Rat); 102137149 (Cynomolgus Monkey)
Alternate Names
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

Citations for Recombinant Mouse Lipocalin-2/NGAL Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

19 Citations: Showing 1 - 10
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  1. Role of lipocalin-2 in surgery-induced cognitive decline in mice: a signal from neuron to microglia
    Authors: X Xiang, X Tang, Y Yu, S Xie, L Liu, M Chen, R Zhang, X Kang, Y Zheng, G Yang, S Gan, S Zhu
    Journal of Neuroinflammation, 2022;19(1):92.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo
  2. Critical changes in hypothalamic gene networks in response to pancreatic cancer as found by single-cell RNA sequencing
    Authors: C Huisman, MA Norgard, PR Levasseur, SM Krasnow, MGP van der Wi, B Olson, DL Marks
    Molecular Metabolism, 2022;0(0):101441.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo
  3. Photobiomodulation inhibits the activation of neurotoxic microglia and astrocytes by inhibiting Lcn2/JAK2-STAT3 crosstalk after spinal cord injury in male rats
    Authors: 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
    Journal of Neuroinflammation, 2021;18(1):256.
    Species: Mouse/Rat
    Sample Types: Whole Cell
    Applications: Bioassay
  4. Electroacupuncture improves repeated social defeat stress-elicited social avoidance and anxiety-like behaviors by reducing Lipocalin-2 in the hippocampus
    Authors: YH Chen, SY Xie, CW Chen, DY Lu
    Molecular Brain, 2021;14(1):150.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo
  5. Chronic cerebral lipocalin 2 exposure elicits hippocampal neuronal dysfunction and cognitive impairment
    Authors: B Olson, X Zhu, MA Norgard, P Diba, PR Levasseur, AC Buenafe, C Huisman, KG Burfeind, KA Michaelis, G Kong, T Braun, DL Marks
    Brain, Behavior, and Immunity, 2021;0(0):.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo
  6. Lipocalin 2 mediates appetite suppression during pancreatic cancer cachexia
    Authors: 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
    Nature Communications, 2021;12(1):2057.
    Species: Mouse
    Sample Types: In Vivo
    Applications: Bioassay
  7. T Cell-Intrinsic IRF5 Regulates T Cell Signaling, Migration, and Differentiation and Promotes Intestinal Inflammation
    Authors: J Yan, SP Pandey, BJ Barnes, JR Turner, C Abraham
    Cell Rep, 2020;31(13):107820.
    Species: Mouse
    Sample Types: Reference Standard
  8. Lipocalin-2 (Lcn-2) Attenuates Polymicrobial Sepsis with LPS Preconditioning (LPS Tolerance) in FcGRIIb Deficient Lupus Mice
    Authors: T Ondee, J Gillen, P Visitchana, P Somparn, J Issara-Amp, C Dang Phi, W Chancharoe, D Gurusamy, A Nita-Lazar, A Leelahavan
    Cells, 2019;8(9):.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo
  9. Lipocalin-2 Regulates Epidermal Growth Factor Receptor Intracellular Trafficking
    Authors: L Yammine, A Zablocki, W Baron, F Terzi, M Gallazzini
    Cell Rep, 2019;29(7):2067-2077.e6.
    Species: Human
    Sample Types: Whole Cell
    Applications: Cell Culture
  10. Lipocalin-2 regulates adult neurogenesis and contextual discriminative behaviours
    Authors: 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
    Mol. Psychiatry, 2017;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  11. Obesity-promoting and anti-thermogenic effects of neutrophil gelatinase-associated lipocalin in mice
    Authors: A Ishii, G Katsuura, H Imamaki, H Kimura, KP Mori, T Kuwabara, M Kasahara, H Yokoi, K Ohinata, T Kawanishi, J Tsuchida, Y Nakamoto, K Nakao, M Yanagita, M Mukoyama, K Mori
    Sci Rep, 2017;7(1):15501.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo
  12. The adipokine lipocalin-2 in the context of the osteoarthritic osteochondral junction
    Sci Rep, 2016;6(0):29243.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  13. Lipocalin 2: a new mechanoresponding gene regulating bone homeostasis.
    Authors: Rucci N, Capulli M, Piperni S, Cappariello A, Lau P, Frings-Meuthen P, Heer M, Teti A
    J Bone Miner Res, 2015;30(2):357-68.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  14. Tcf3 promotes cell migration and wound repair through regulation of lipocalin 2.
    Authors: Miao Q, Ku A, Nishino Y, Howard J, Rao A, Shaver T, Garcia G, Le D, Karlin K, Westbrook T, Poli V, Nguyen H
    Nat Commun, 2014;5(0):4088.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo
  15. CCR5 knockout prevents neuronal injury and behavioral impairment induced in a transgenic mouse model by a CXCR4-using HIV-1 glycoprotein 120.
    Authors: 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
    J Immunol, 2014;193(4):1895-910.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  16. Differential role of lipocalin 2 during immune complex-mediated acute and chronic inflammation in mice.
    Authors: Shashidharamurthy R, Machiah D, Aitken J, Putty K, Srinivasan G, Chassaing B, Parkos C, Selvaraj P, Vijay-Kumar M
    Arthritis Rheum, 2013;65(4):1064-73.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo
  17. Role of lipocalin-2-chemokine axis in the development of neuropathic pain following peripheral nerve injury.
    Authors: Jeon, Sangmin, Jha, Mithiles, Ock, Jiyeon, Seo, Jungwan, Jin, Myungwon, Cho, Heejung, Lee, Won-Ha, Suk, Kyoungho
    J Biol Chem, 2013;288(33):24116-27.
    Species: Mouse
    Sample Types: In Vivo
    Applications: In Vivo
  18. Absence of functional Hfe protects mice from invasive Salmonella enterica serovar Typhimurium infection via induction of lipocalin-2.
    Authors: Nairz M, Theurl I, Schroll A, Theurl M, Fritsche G, Lindner E, Seifert M, Crouch ML, Hantke K, Akira S, Fang FC, Weiss G
    Blood, 2009;114(17):3642-51.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  19. The role of lipocalin 2 in the regulation of inflammation in adipocytes and macrophages.
    Authors: Zhang J, Wu Y, Zhang Y, LeRoith D, Bernlohr DA, Chen X
    Mol. Endocrinol., 2008;22(6):1416-26.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay

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Recombinant Mouse Lipocalin-2/NGAL Protein, CF
By Vishal Singh on 05/23/2016
Application: Iron and bacterial siderophore binding capacity
Reason for Rating: Carrier and siderophore free murine Lcn2 helps us to analyze the its siderophore and iron binding capacity in in vitro test systems.