Human MMP-9/NGAL Complex Quantikine ELISA Kit

Newer Version Available: DY8556-05
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Human MMP-9/Lipocalin-2 Complex ELISA Standard Curve
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Citations (15)
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Human MMP-9/NGAL Complex Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (40 uL), Serum (10 uL), Heparin Plasma (10 uL), Saliva (10 uL), Urine (40 uL)
Sensitivity
0.058 ng/mL
Assay Range
0.3 - 20 ng/mL (Cell Culture Supernates, Serum, Heparin Plasma, Saliva, Urine)
Specificity
Natural human MMP-9/NGAL Complex
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
Interference observed with 1 or more available related molecules.

Product Summary

The Quantikine Human MMP-9/NGAL Complex Immunoassay is a 4.5 hour solid phase ELISA designed to measure the MMP-9/NGAL complex in cell culture supernates, serum, plasma, urine, and saliva. It contains natural human MMP-9/NGAL as the standard. The antibodies were raised against recombinant human MMP-9 and recombinant human NGAL. This kit will not detect recombinant human MMP-9 or NGAL in their free forms.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, Heparin Plasma, Saliva, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 1.84 5.52 10.3 1.7 5.13 9.92
Standard Deviation 0.076 0.16 0.237 0.13 0.335 0.51
CV% 4.1 2.9 2.3 7.6 6.5 5.1

Recovery

The recovery of natural human MMP-9/NGAL Complex spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 90 84-97
Urine (n=4) 93 86-106

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of natural MMP-9/NGAL Complex were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human MMP-9/Lipocalin-2 Complex ELISA Linearity

Scientific Data

Human MMP-9/Lipocalin-2 Complex ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
&#9888; WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human MMP-9/NGAL Complex Quantikine ELISA Kit

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.

15 Citations: Showing 1 - 10
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  1. Urinary NGAL Measured after the First Year Post Kidney Transplantation Predicts Changes in Glomerular Filtration over One-Year Follow-Up
    Authors: M Kielar, P Dumnicka, A Gala-B??dz, A B?dkowska-, E Ignacak, B Maziarz, P Ceranowicz, B Ku?nierz-C
    Journal of Clinical Medicine, 2020-12-25;10(1):.
    Species: Human
    Sample Types: Urine
  2. Determination of FABP4, RBP4 and the MMP-9/NGAL complex in the serum of women with breast cancer
    Authors: D Tsakogiann, E Kalogera, F Zagouri, E Zografos, D Balalis, G Bletsa
    Oncology Letters, 2020-12-04;21(2):85.
    Species: Human
    Sample Types: Serum
  3. Lipocalin 2 as a potential systemic biomarker for central serous chorioretinopathy
    Authors: A Matet, T Jaworski, E Bousquet, J Canonica, C Gobeaux, A Daruich, M Zhao, M Zola, M Meester-Sm, D Mohabati, F Jaisser, S Yzer, F Behar-Cohe
    Sci Rep, 2020-11-19;10(1):20175.
    Species: Human
    Sample Types: Serum
  4. Changes in brain white matter structure are associated with urine proteins in urologic chronic pelvic pain syndrome (UCPPS): A MAPP Network study
    Authors: DC Woodworth, A Dagher, A Curatolo, M Sachdev, C Ashe-McNal, BD Naliboff, JS Labus, JR Landis, JJ Kutch, EA Mayer, RS Lee, MA Moses, BM Ellingson
    PLoS ONE, 2018-12-05;13(12):e0206807.
    Species: Human
    Sample Types: Urine
  5. Neutrophil gelatinase associated lipocalin (NGAL) is elevated in type 2 diabetics with carotid artery stenosis and reduced under metformin treatment
    Authors: W Eilenberg, S Stojkovic, A Piechota-P, A Kaider, N Kozakowski, WJ Weninger, J Nanobachvi, J Wojta, I Huk, S Demyanets, C Neumayer
    Cardiovasc Diabetol, 2017-08-08;16(1):98.
    Species: Human
    Sample Types: Serum
  6. A case-control study of pre-operative levels of serum neutrophil gelatinase-associated lipocalin and other potential inflammatory markers in colorectal cancer.
    Authors: Duvillard L, Ortega-Deballon P, Bourredjem A, Scherrer M, Mantion G, Delhorme J, Deguelte-Lardiere S, Petit J, Bonithon-Kopp C
    BMC Cancer, 2014-12-03;14(0):912.
    Species: Human
    Sample Types: Serum
  7. Molecular MRI of murine atherosclerotic plaque targeting NGAL: a protein associated with unstable human plaque characteristics.
    Authors: te Boekhorst BC, Bovens SM, Hellings WE, van der Kraak PH, van de Kolk KW, Vink A, Moll FL, van Oosterhout MF, de Vries JP, Doevendans PA, Goumans MJ, de Kleijn DP, van Echteld CJ, Pasterkamp G, Sluijter JP
    Cardiovasc. Res., 2010-10-28;89(3):680-8.
    Species: Human
    Sample Types: Plasma
  8. Primary human acute myelogenous leukemia cells release matrix metalloproteases and their inhibitors: release profile and pharmacological modulation.
    Authors: Reikvam H, Hatfield KJ, Oyan AM, Kalland KH, Kittang AO, Bruserud O
    Eur. J. Haematol., 2009-11-17;84(3):239-51.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Circulating levels of matrix metalloproteinase-9 (MMP-9), neutrophil gelatinase-associated lipocalin (NGAL) and their complex MMP-9/NGAL in breast cancer disease.
    Authors: Provatopoulou X, Gounaris A, Kalogera E
    BMC Cancer, 2009-11-04;9(0):390.
    Species: Human
    Sample Types: Serum
  10. Differential inflammatory activity across human abdominal aortic aneurysms reveals neutrophil-derived leukotriene B4 as a major chemotactic factor released from the intraluminal thrombus.
    Authors: Houard X, Ollivier V, Louedec L, Michel JB, Back M
    FASEB J., 2009-01-09;23(5):1376-83.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Urinary biomarkers predict brain tumor presence and response to therapy.
    Authors: Smith ER, Zurakowski D, Saad A, Scott RM, Moses MA
    Clin. Cancer Res., 2008-04-15;14(8):2378-86.
    Species: Human
    Sample Types: Urine
  12. Serum concentrations of atherogenic proteins neutrophil gelatinase-associated lipocalin and its complex with matrix metalloproteinase-9 are significantly lower in women with polycystic ovary syndrome: hint of a protective mechanism?
    Authors: Diamanti-Kandarakis E, Livadas S, Kandarakis SA, Margeli A, Papassotiriou I
    Eur. J. Endocrinol., 2008-04-01;158(4):525-31.
    Species: Human
    Sample Types: Serum
  13. Increased expression of discoidin domain receptor 2 is linked to the degree of cartilage damage in human knee joints: a potential role in osteoarthritis pathogenesis.
    Authors: Sunk IG, Bobacz K, Hofstaetter JG, Amoyo L, Soleiman A, Smolen J, Xu L, Li Y
    Arthritis Rheum., 2007-11-01;56(11):3685-92.
    Species: Human
    Sample Types: Plasma
  14. Presence of NGAL/MMP-9 complexes in human abdominal aortic aneurysms.
    Authors: Folkesson M, Kazi M, Zhu C, Silveira A, Hemdahl AL, Hamsten A, Hedin U, Swedenborg J, Eriksson P
    Thromb. Haemost., 2007-08-01;98(2):427-33.
    Species: Human
    Sample Types: Tissue Homogenates
  15. Low plasma levels of HSP70 in patients with carotid atherosclerosis are associated with increased levels of proteolytic markers of neutrophil activation.
    Authors: Martin-Ventura JL, Leclercq A, Blanco-Colio LM, Egido J, Rossignol P, Meilhac O, Michel JB
    Atherosclerosis, 2006-12-08;194(2):334-41.
    Species: Human
    Sample Types: Plasma

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