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Recombinant Human Lipocalin-2/NGAL Protein, CF

Catalog #: 1757-LC
23 Citations 1 Review Datasheet
Catalog # Availability Size / Price Qty
1757-LC-050
R&D Systems Recombinant Proteins and Enzymes
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Citations (23)
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Reviews (1)
Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Lipocalin-2/NGAL Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
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. 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 #
P80188
N-terminal Sequence
Analysis
No results obtained: Gln21 predicted
Predicted Molecular Mass
22 kDa
SDS-PAGE
25-27 kDa doublet, reducing conditions

Product Datasheets

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1757-LC

Product Datasheet

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.

1757-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)
  • 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

Background: Lipocalin-2/NGAL

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).

References
  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.
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 Human 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.

23 Citations: Showing 1 - 10
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  1. Mycobiota and diet-derived fungal xenosiderophores promote Salmonella gastrointestinal colonization
    Authors: W Santus, AP Rana, JR Devlin, KA Kiernan, CC Jacob, J Tjokrosurj, DM Underhill, J Behnsen
    Nature Microbiology, 2022-11-21;7(12):2025-2038.
    Species: Salmonella
    Sample Types: Whole Cells
    Applications: Bioassay
  2. Increased LCN2 (lipocalin 2) in the RPE decreases autophagy and activates inflammasome-ferroptosis processes in a mouse model of dry AMD
    Authors: 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
    Autophagy, 2022-04-26;0(0):1-20.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  3. ADAM8-Dependent Extracellular Signaling in the Tumor Microenvironment Involves Regulated Release of Lipocalin 2 and MMP-9
    Authors: 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
    International Journal of Molecular Sciences, 2022-02-10;23(4):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Stimulation
  4. Lipocalin-2 (LCN2) Deficiency Leads to Cellular Changes in Highly Metastatic Human Prostate Cancer Cell Line PC-3
    Authors: SK Schröder, M Pinoé-Schm, R Weiskirche
    Cells, 2022-01-13;11(2):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  5. Epigenetic induction of lipocalin 2 expression drives acquired resistance to 5-fluorouracil in colorectal cancer through integrin beta3/SRC pathway
    Authors: 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
    Oncogene, 2021-09-29;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Bioassay
  6. Obesity-associated up-regulation of lipocalin 2 protects gastric mucosa cells from apoptotic cell death by reducing endoplasmic reticulum stress
    Authors: 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
    Cell Death & Disease, 2021-02-26;12(2):221.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  7. Nicotine promotes breast cancer metastasis by stimulating N2 neutrophils and generating pre-metastatic niche in lung
    Authors: A Tyagi, S Sharma, K Wu, SY Wu, F Xing, Y Liu, D Zhao, RP Deshpande, RB D'Agostino, K Watabe
    Nature Communications, 2021-01-20;12(1):474.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  8. Increased expression of LCN2 formed a positive feedback loop with activation of the ERK pathway in human kidney cells during kidney stone formation
    Authors: Z Hui, Z Jiang, D Qiao, Z Bo, K Qiyuan, B Shaohua, Y Wenbing, L Wei, L Cheng, L Shuangning, L Zhengyi, L Yingyi
    Scientific Reports, 2020-12-04;10(1):21287.
    Species: Human
    Sample Types: Whole Cells
    Applications: Cell Culture
  9. Subtilase cytotoxin induces a novel form of Lipocalin 2, which promotes Shiga-toxigenic Escherichia coli survival
    Authors: K Yahiro, K Ogura, Y Goto, S Iyoda, T Kobayashi, H Takeuchi, M Ohnishi, J Moss
    Sci Rep, 2020-11-03;10(1):18943.
    Species: Human
    Sample Types: Whole Cells
    Applications: Cell Culture
  10. Relation of Neutrophil Gelatinase-Associated Lipocalin Overexpression to the Resistance to Apoptosis of Tumor B Cells in Chronic Lymphocytic Leukemia
    Authors: B Bauvois, E Pramil, L Jondrevill, E Chapiro, C Quiney, K Maloum, SA Susin, F Nguyen-Kha
    Cancers (Basel), 2020-07-31;12(8):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  11. Neutrophil extracellular traps (NETs) contribute to pathological changes of ocular graft-vs.-host disease (oGVHD) dry eye: Implications for novel biomarkers and therapeutic strategies
    Authors: S An, I Raju, B Surenkhuu, JE Kwon, S Gulati, M Karaman, A Pradeep, S Sinha, C Mun, S Jain
    Ocul Surf, 2019-04-06;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  12. TCF7L2 and EGR1 synergistic activation of transcription of LCN2 via an ERK1/2-dependent pathway in esophageal squamous cell carcinoma cells
    Authors: Y Zhao, Q Xia, Y Liu, W Bai, Y Yao, J Ding, L Lin, Z Xu, Z Cai, S Wang, E Li, H Xu, B Wu, L Xu, Z Du
    Cell. Signal., 2018-12-14;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  13. Lipocalin 2 Plays an Important Role in Regulating Inflammation in Retinal Degeneration
    Authors: T Parmar, VM Parmar, L Perusek, A Georges, M Takahashi, JW Crabb, A Maeda
    J. Immunol., 2018-03-30;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  14. Neutrophil Gelatinase-Associated Lipocalin Attenuates Ischemia/Reperfusion Injury in an In Vitro Model via Autophagy Activation
    Authors: C Yan, T Yuanjie, X Zhengqun, C Jiayan, L Kongdan
    Med. Sci. Monit., 2018-01-25;24(0):479-485.
    Species: Rat
    Sample Types: Whole Cells
    Applications: Bioassay
  15. Lipocalin-2 Promotes Pancreatic Ductal Adenocarcinoma by Regulating Inflammation in the Tumor Microenvironment
    Authors: S Gomez-Chou, A Swidnicka-, N Badi, M Chavez-Tom, GB Lesinski, T Bekaii-Saa, MR Farren, TA Mace, C Schmidt, Y Liu, D Deng, RF Hwang, L Zhou, TT Moore, D Chatterjee, H Wang, X Leng, RB Arlinghaus, CD Logsdon, Z Cruz-Monse
    Cancer Res, 2017-03-01;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  16. Neutrophil gelatinase-associated lipocalin predicts myocardial dysfunction and mortality in severe sepsis and septic shock
    Authors: Xiaoye Yan
    Int. J. Cardiol., 2016-11-03;0(0):.
    Species: Human
    Sample Types: Plasma
    Applications: ELISA Developmet
  17. Gut Microbiota Conversion of Dietary Ellagic Acid into Bioactive Phytoceutical Urolithin A Inhibits Heme Peroxidases
    PLoS ONE, 2016-06-02;11(6):e0156811.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  18. Neutrophil Gelatinase Associated Lipocalin Is an Early and Accurate Biomarker of Graft Function and Tissue Regeneration in Kidney Transplantation from Extended Criteria Donors.
    Authors: Cantaluppi V, Dellepiane S, Tamagnone M, Medica D, Figliolini F, Messina M, Manzione A, Gai M, Tognarelli G, Ranghino A, Dolla C, Ferrario S, Tetta C, Segoloni G, Camussi G, Biancone L
    PLoS ONE, 2015-06-30;10(6):e0129279.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  19. Estrogen receptor (ER)alpha-regulated lipocalin 2 expression in adipose tissue links obesity with breast cancer progression.
    Authors: Drew B, Hamidi H, Zhou Z, Villanueva C, Krum S, Calkin A, Parks B, Ribas V, Kalajian N, Phun J, Daraei P, Christofk H, Hewitt S, Korach K, Tontonoz P, Lusis A, Slamon D, Hurvitz S, Hevener A
    J Biol Chem, 2014-12-02;290(9):5566-81.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  20. Neutrophil gelatinase-associated lipocalin increases HLA-G(+)/FoxP3(+) T-regulatory cell population in an in vitro model of PBMC.
    Authors: La Manna G, Ghinatti G, Tazzari P, Alviano F, Ricci F, Capelli I, Cuna V, Todeschini P, Brunocilla E, Pagliaro P, Bonsi L, Stefoni S
    PLoS ONE, 2014-02-27;9(2):e89497.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  21. Lipocalin-2 increases fat oxidation in vitro and is correlated with energy expenditure in normal weight but not obese women.
    Authors: Paton C, Rogowski M, Kozimor A, Stevenson J, Chang H, Cooper J
    Obesity (Silver Spring), 2013-07-29;21(12):E640-8.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Cell Culture
  22. Lipocalin 2 regulation by thermal stresses: protective role of Lcn2/NGAL against cold and heat stresses.
    Authors: Roudkenar MH, Halabian R, Roushandeh AM, Nourani MR, Masroori N, Ebrahimi M, Nikogoftar M, Rouhbakhsh M, Bahmani P, Najafabadi AJ, Shokrgozar MA
    Exp. Cell Res., 2009-09-02;315(18):3140-51.
    Species: Hamster, Human
    Sample Types: Whole Cells
    Applications: Bioassay
  23. Urinary lipocalin-2 is associated with renal disease activity in human lupus nephritis.
    Authors: Pitashny M, Schwartz N, Qing X, Hojaili B, Aranow C, Mackay M, Putterman C
    Arthritis Rheum., 2007-06-01;56(6):1894-903.
    Applications: ELISA (Standard), Western Blot

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Recombinant Human Lipocalin-2/NGAL Protein, CF
By Balaji Mahender on 02/05/2018
Application: Immunoassay Standard