Rat Lipocalin-2/NGAL DuoSet ELISA

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Rat Lipocalin-2 / NGAL ELISA Standard Curve
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Product Details
Citations (17)
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Rat Lipocalin-2/NGAL DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Assay Range
78.1 - 5,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant rat Lipocalin-2/NGAL. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

Rat Lipocalin-2 / NGAL ELISA Standard Curve

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Lipocalin-2/NGAL

The Lipocalin family comprises a diverse group of mostly secreted soluble proteins that bind hydrophobic ligands and act as transporters, carrying small molecules to specific cells. Lipocalins are related by possessing an 8-stranded beta-barrel structure. Lipocalin-1, also named tear lipocalin (TL), von Ebners gland protein (VEG) and tear pre-albumin, binds a large number of hydrophobic molecules and exhibits cysteine proteinase inhibitor and endonuclear activities. Lipocalin-2, also known as neutrophil gelatinase-associated lipocalin (NGAL), is a component of granules in neutrophils from tissues that are normally exposed to microorganisms and is upregulated during inflammation. Lipocalin-2 can form homodimers and can heterodimerize with the neutrophil gelatinase MMP-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

Assay Procedure


Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Rat Lipocalin-2/NGAL DuoSet ELISA

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.

17 Citations: Showing 1 - 10
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  1. Effects of a lysophosphatidic acid receptor 1 antagonist on hypertensive renal injury in Dahl-Iwai salt-sensitive rats
    Authors: T Naruse, H Otake, T Takahashi
    Oncogene, 2022;149(4):179-188.
    Species: Rat
    Sample Types: Urine
  2. miR?214 ameliorates sepsis?induced acute kidney injury via PTEN/AKT/mTOR?regulated autophagy
    Authors: Z Sang, S Dong, P Zhang, Y Wei
    Molecular Medicine Reports, 2021;24(4):.
    Species: Mouse
    Sample Types: Urine
  3. Study of the Role of the Tyrosine Kinase Receptor MerTK in the Development of Kidney Ischemia-Reperfusion Injury in RCS Rats
    Authors: T Pelé, S Giraud, S Joffrion, V Ameteau, A Delwail, JM Goujon, L Macchi, T Hauet, F Dkhissi, O Benzakour
    International Journal of Molecular Sciences, 2021;22(22):.
    Species: Rat
    Sample Types: Plasma
  4. Fecal dysbiosis associated with colonic hypersensitivity and behavioral alterations in chronically Blastocystis-infected rats
    Authors: M Defaye, C Nourrisson, E Baudu, A Lashermes, M Meynier, M Meleine, I Wawrzyniak, V Bonnin, J Barbier, B Chassaing, C Godfraind, A Gelot, N Barnich, D Ardid, M Bonnet, F Delbac, FA Carvalho, P Poirier
    Sci Rep, 2020;10(1):9146.
    Species: Rat
    Sample Types: Tissue Homogenates
  5. Rats with congenital hydronephrosis show increased susceptibility to renal ischemia-reperfusion injury
    Authors: R Vilskersts, K Vilks, M Videja, H Cirule, O Zharkova-M, E Sevostjano, M Dambrova, E Liepinsh
    Physiol Rep, 2020;8(22):e14638.
    Species: Rat
    Sample Types: Urine
  6. Glomerular Collagen Deposition and Lipocalin-2 Expression Are Early Signs of Renal Injury in Prediabetic Obese Rats
    Authors: EN Bukosza, T Kaucsár, M Godó, E Lajtár, P Tod, G Koncsos, ZV Varga, T Baranyai, NM Tu, H Schachner, C S?ti, P Ferdinandy, Z Giricz, G Szénási, P Hamar
    Int J Mol Sci, 2019;20(17):.
    Species: Rat
    Sample Types: Plasma
  7. Hypoxia Induces Astrocyte-Derived Lipocalin-2 in Ischemic Stroke
    Authors: F Ranjbar Ta, N Gasterich, M Scheld, R Weiskirche, C Beyer, T Clarner, A Zendedel
    Int J Mol Sci, 2019;20(6):.
  8. Renal injury after uninephrectomy in male and female intrauterine growth-restricted aged rats
    Authors: AD Newsome, GK Davis, ON Adah, NB Ojeda, BT Alexander
    PLoS ONE, 2019;14(3):e0213404.
    Species: Rat
    Sample Types: Urine
  9. TGF-?-mediated NADPH oxidase 4-dependent oxidative stress promotes colistin-induced acute kidney injury
    Authors: BY Jeong, SR Park, S Cho, SL Yu, HY Lee, CG Park, J Kang, DY Jung, MH Park, WM Hwang, SR Yun, JY Jung, SH Yoon
    J. Antimicrob. Chemother., 2018;0(0):.
    Species: Rat
    Sample Types: Serum
  10. Salt-Responsive Metabolite, ?-Hydroxybutyrate, Attenuates Hypertension
    Authors: S Chakrabort, S Galla, X Cheng, JY Yeo, B Mell, V Singh, B Yeoh, P Saha, AV Mathew, M Vijay-Kuma, B Joe
    Cell Rep, 2018;25(3):677-689.e4.
    Species: Rat
    Sample Types: Serum
  11. Neutrophil gelatinase-associated lipocalin level is a prognostic factor for survival in rat and human chronic liver diseases
    Authors: K Yoshikawa, M Iwasa, A Eguchi, S Kojima, N Yoshizawa, M Tempaku, R Sugimoto, N Yamamoto, K Sugimoto, Y Kobayashi, H Hasegawa, Y Takei
    Hepatol Commun, 2017;1(9):946-956.
    Species: Rat
    Sample Types: Plasma
    Applications: ELISA Capture
  12. Insulin treatment prevents wounding associated changes in tissue and circulating neutrophil MMP-9 and NGAL in diabetic rats
    Authors: M Abdollahi, TS Ng, A Rezaeizade, S Aamidor, SM Twigg, D Min, SV McLennan
    PLoS ONE, 2017;12(2):e0170951.
    Species: Rat
    Sample Types: Tissue Secretion
  13. Cardioprotection and lifespan extension by the natural polyamine spermidine
    Nat. Med., 2016;0(0):.
    Species: Rat
    Sample Types: Urine
  14. Chronic recurrent dehydration associated with periodic water intake exacerbates hypertension and promotes renal damage in male spontaneously hypertensive rats
    Sci Rep, 2016;6(0):33855.
    Species: Rat
    Sample Types: Urine
  15. Oxidative/Nitrative Stress and Inflammation Drive Progression of Doxorubicin-Induced Renal Fibrosis in Rats as Revealed by Comparing a Normal and a Fibrosis-Resistant Rat Strain.
    Authors: Szalay C, Erdelyi K, Kokeny G, Lajtar E, Godo M, Revesz C, Kaucsar T, Kiss N, Sarkozy M, Csont T, Krenacs T, Szenasi G, Pacher P, Hamar P
    PLoS ONE, 2015;10(6):e0127090.
    Species: Rat
    Sample Types: Urine
  16. Modulation of early inflammatory response by different balanced and non-balanced colloids and crystalloids in a rodent model of endotoxemia.
    Authors: Voigtsberger S, Urner M, Hasler M, Roth Z'Graggen B, Booy C, Spahn D, Beck-Schimmer B
    PLoS ONE, 2014;9(4):e93863.
    Species: Rat
    Sample Types: Urine
  17. Intrarenal resistance index for the assessment of acute renal injury in a rat liver transplantation model.
    Authors: Kong H, Chen F, He Y, Wu L, Wang L, Zhu S, Zheng S
    BMC Nephrol, 2013;14(0):55.
    Species: Rat
    Sample Types: Serum


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