Human TIMP-2 Quantikine ELISA Kit

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DTM200
Control Products Available
Human TIMP-2 ELISA Cell Culture Supernate/Saliva/Urine Standard Curve
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Product Details
Procedure
Citations (36)
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Human TIMP-2 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 (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Saliva (10 uL), Urine (25 uL)
Sensitivity
0.064 ng/mL
Assay Range
0.2 - 10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
Specificity
Natural and recombinant human TIMP-2. This assay detects approximately 50% of recombinant human TIMP-2 when complexed with recombinant human active MMP-9 in a 1:1 molar ratio.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human TIMP-2 Immunoassay is a 4.5 hour solid phase ELISA designed to measure TIMP-2 levels in cell culture supernates, serum, plasma, and saliva. It contains CHO cell-expressed recombinant human TIMP-2, and antibodies raised against the recombinant protein. Natural human TIMP-2 showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards, indicating that this kit can be used to determine relative levels of natural human TIMP-2.

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, Saliva

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 1.01 2.9 5.26 1.01 2.79 5.27
Standard Deviation 0.042 0.173 0.344 0.079 0.188 0.326
CV% 4.2 6 6.5 7.8 6.7 6.2

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 1.23 3.45 6.09 1.26 3.45 6.38
Standard Deviation 0.054 0.116 0.181 0.086 0.197 0.467
CV% 4.4 3.4 3 6.8 5.7 7.3

Recovery

The recovery of TIMP-2 spiked to three different levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 95 81-108

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of TIMP-2 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human TIMP-2 Linearity

Scientific Data

Human TIMP-2 ELISA Cell Culture Supernate/Saliva/Urine Standard Curve

Human TIMP-2 ELISA Serum/Plasma 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.

Background: TIMP-2

TIMPs-1 through -4 regulate the activity of zinc metalloproteases known as MMPs, ADAMs and ADAMTSs. Structurally, TIMPs contain two domains. The N-terminal domain binds to the active site of mature metalloproteases via a 1:1 non-covalent interaction, blocking access of substrates to the catalytic site. In addition, The C-terminal domain of TIMP-1 and TIMP-2 binds to the hemopexin- like domain of pro-MMP-9 and pro-MMP-2, respectively. The latter binding is essential for the cell surface activation of MMP-2 by MMP-14.

Long Name:
Tissue Inhibitors of Metalloproteinases 2
Entrez Gene IDs:
7077 (Human); 21858 (Mouse); 29543 (Rat)
Alternate Names:
CSC-21Ktissue inhibitor of metalloproteinase 2; metalloproteinase inhibitor 2; TIMP metallopeptidase inhibitor 2; TIMP2; TIMP-2; Tissue inhibitor of metalloproteinases 2
&#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 2 hours on a horizontal orbital microplate shaker.
  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 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well.
  13.   For Cell Culture Supernate & Saliva Samples: Incubate at room temperature for 20 minutes on the benchtop. PROTECT FROM LIGHT.
    For Serum & Plasma Samples: Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  14. 50 µL Stop Solution
  15. 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 TIMP-2 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.

36 Citations: Showing 1 - 10
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  1. Urinary beta-2 microglobulin increases whereas TIMP-2 and IGFBP7 decline after unilateral nephrectomy in healthy kidney donors
    Authors: de Rooij, ENM;Hoogeveen, EK;Romijn, FPHTM;van der Kooy, SW;Veighey, KV;Dekker, FW;van Kooten, C;Cobbaert, CM;de Fijter, JW;
    Scientific reports
    Species: Human
    Sample Types: Urine
  2. Enzymes of Fibrosis in Chronic Liver Disease
    Authors: I Tsomidis, G Notas, C Xidakis, A Voumvourak, DN Samonakis, M Koulentaki, E Kouroumali
    Biomedicines, 2022-12-08;10(12):.
    Species: Human
    Sample Types: Serum
  3. Estrogen Receptor Subtypes Elicit a Distinct Gene Expression Profile of Endothelial-Derived Factors Implicated in Atherosclerotic Plaque Vulnerability
    Authors: N Nasiri-Ans, E Spilioti, I Kyrou, V Kalotychou, A Chatzigeor, D Sanoudou, K Dahlman-Wr, HS Randeva, AG Papavassil, P Moutsatsou, E Kassi
    International Journal of Molecular Sciences, 2022-09-19;23(18):.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Circulating biomarker correlates of left atrial size and myocardial extracellular volume fraction among persons living with and without HIV
    Authors: TE Peterson, C Landon, SA Haberlen, F Bhondoekha, MW Plankey, FJ Palella, DA Piggott, JB Margolick, TT Brown, WS Post, KC Wu
    BMC cardiovascular disorders, 2022-09-03;22(1):393.
    Species: Human
    Sample Types: Serum
  5. The ASIC3-M-CSF-M2 macrophage-positive feedback loop modulates fibroblast-to-myofibroblast differentiation in skin fibrosis pathogenesis
    Authors: JJ Wu, ZL Sun, SY Liu, ZH Chen, ZD Yuan, ML Zou, YY Teng, YY Li, DY Guo, FL Yuan
    Cell Death & Disease, 2022-06-06;13(6):527.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Combining renal cell arrest and damage biomarkers to predict progressive AKI in patient with sepsis
    Authors: X Tao, C Chen, W Luo, J Zhou, J Tian, X Yang, FF Hou
    Bmc Nephrology, 2021-12-15;22(1):415.
    Species: Human
    Sample Types: Urine
  7. Zinc Administration and Improved Serum Markers of Hepatic Fibrosis in Patients with Autoimmune Hepatitis
    Authors: K Moriya, N Nishimura, T Namisaki, H Takaya, Y Sawada, H Kawaratani, K Kaji, N Shimozato, S Sato, M Furukawa, A Douhara, T Akahane, A Mitoro, J Yamao, H Yoshiji
    Journal of Clinical Medicine, 2021-06-02;10(11):.
    Species: Human
    Sample Types: Serum
  8. The predictive value of TIMP-2 and IGFBP7 for kidney failure and 30-day mortality after elective cardiac surgery
    Authors: K Esmeijer, A Schoe, LR Ruhaak, EK Hoogeveen, D Soonawala, FPHTM Romijn, MR Shirzada, JT van Dissel, CM Cobbaert, JW de Fijter
    Scientific Reports, 2021-01-13;11(1):1071.
    Species: Human
    Sample Types: Urine
  9. Matrix Metalloproteinases MMP-2 and MMP-9, Their Inhibitors TIMP-1 and TIMP-2, Vascular Endothelial Growth Factor and sVEGFR-2 as Predictive Markers of Ischemic Retinopathy in Patients with Systemic Sclerosis-Case Series Report
    Authors: A Waszczykow, M Podgórski, M Waszczykow, Z Gerlicz-Ko, P Jurowski
    Int J Mol Sci, 2020-11-18;21(22):.
    Species: Human
    Sample Types: Serum
  10. Cdc42-Dependent Transfer of mir301 from Breast Cancer-Derived Extracellular Vesicles Regulates the Matrix Modulating Ability of Astrocytes at the Blood-Brain Barrier
    Authors: G Morad, CC Daisy, HH Otu, TA Libermann, ST Dillon, MA Moses
    Int J Mol Sci, 2020-05-28;21(11):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  11. Serum melatonin is inversely associated with matrix metalloproteinase-9 in oral squamous cell carcinoma
    Authors: AE Stanciu, A Zamfir-Chi, MM Stanciu, A Pantea-Sto, C Nitipir, DC Gheorghe
    Oncol Lett, 2020-02-13;19(4):3011-3020.
    Species: Human
    Sample Types: Serum
  12. Levels of Selected Matrix Metalloproteinases, Their Inhibitors in Saliva, and Oral Status in Juvenile Idiopathic Arthritis Patients vs. Healthy Controls
    Authors: A Kobus, J Bagi?ska, J ?api?ska-A, S ?awicki, A Kierklo
    Biomed Res Int, 2019-10-28;2019(0):7420345.
    Species: Human
    Sample Types: Saliva
  13. Basilar Artery Tortuosity Is Associated With White Matter Hyperintensities by TIMP-1
    Authors: DP Zhang, YF Peng, HL Zhang, JG Ma, M Zhao, S Yin, TT Wei
    Front Neurosci, 2019-08-14;13(0):836.
    Species: Human
    Sample Types: Plasma
  14. Urinary Tissue Inhibitor of�Metalloproteinases-2 and Insulin-Like Growth Factor-Binding Protein 7 Do Not Correlate With Disease Severity in ADPKD Patients
    Authors: SEI Dekker, LR Ruhaak, FPHTM Romijn, E Meijer, CM Cobbaert, JW de Fijter, D Soonawala
    Kidney Int Rep, 2019-03-22;4(6):833-841.
    Species: Human
    Sample Types: Urine
  15. A potential key mechanism in ascending aortic aneurysm development: Detection of a linear relationship between MMP-14/TIMP-2 ratio and active MMP-2
    Authors: R Schmitt, A Tscheuschl, P Laschinski, X Uffelmann, P Discher, J Fuchs, M Kreibich, R Peyronnet, FA Kari
    PLoS ONE, 2019-02-22;14(2):e0212859.
    Species: Human
    Sample Types: Tissue Homogenates
  16. Matrix metalloproteinase 2 (MMP-2) and its tissue inhibitor 2 (TIMP-2) in pancreatic cancer (PC)
    Authors: M ?ukaszewic, M Gryko, S P?czek, M Szmitkowsk, B K?dra, B Mroczko
    Oncotarget, 2019-01-08;10(3):395-403.
    Species: Human
    Sample Types: Serum
  17. Anti-cancer effect of dung beetle glycosaminoglycans on melanoma
    Authors: MY Ahn, BJ Kim, HJ Kim, JM Jin, HJ Yoon, JS Hwang, KK Park
    BMC Cancer, 2019-01-05;19(1):9.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. Combination of Plasma Biomarkers and Clinical Data for the Detection of Myocardial Fibrosis or Aggravation of Heart Failure Symptoms in Heart Failure with Preserved Ejection Fraction Patients
    Authors: CK Wu, MM Su, YF Wu, JJ Hwang, LY Lin
    J Clin Med, 2018-11-08;7(11):.
    Species: Human
    Sample Types: Plasma
  19. Human umbilical cord plasma proteins revitalize hippocampal function in aged mice
    Authors: JM Castellano, KI Mosher, RJ Abbey, AA McBride, ML James, D Berdnik, JC Shen, B Zou, XS Xie, M Tingle, IV Hinkson, MS Angst, T Wyss-Coray
    Nature, 2017-04-19;544(7651):488-492.
    Species: Human
    Sample Types: Plasma
  20. TIMP1 and MMP9 are predictors of mortality in septic patients in the emergency department and intensive care unit unlike MMP9/TIMP1 ratio: Multivariate model
    Authors: ME Niño, SE Serrano, DC Niño, DM McCosham, ME Cardenas, VP Villareal, M Lopez, A Pazin-Filh, FA Jaimes, F Cunha, R Schulz, D Torres-Due
    PLoS ONE, 2017-02-13;12(2):e0171191.
    Species: Human
    Sample Types: Serum
  21. Role of serum VEGFA, TIMP2, MMP2 and MMP9 in monitoring response to adjuvant radiochemotherapy in patients with primary cervical cancer--results of a companion protocol of the randomized NOGGO-AGO phase III clinical trial.
    Authors: Braicu E, Gasimli K, Richter R, Nassir M, Kummel S, Blohmer J, Yalcinkaya I, Chekerov R, Ignat I, Ionescu A, Mentze M, Fotopoulou C, Pop C, Lichtenegger W, Sehouli J
    Anticancer Res, 2014-01-01;34(1):385-91.
    Species: Human
    Sample Types: Serum
  22. Role of serum concentration of VEGFR1 and TIMP2 on clinical outcome in primary cervical cancer: results of a companion protocol of the randomized, NOGGO-AGO phase III adjuvant trial of simultaneous cisplatin-based radiochemotherapy vs. carboplatin and paclitaxel containing sequential radiotherapy.
    Authors: Braicu E, Fotopoulou C, Chekerov R, Richter R, Blohmer J, Kummel S, Stamatian F, Yalcinkaya I, Mentze M, Lichtenegger W, Sehouli J
    Cytokine, 2013-02-14;61(3):755-8.
    Species: Human
    Sample Types: Serum
  23. Activation of matrix metalloproteinases following anti-Abeta immunotherapy; implications for microhemorrhage occurrence.
    Authors: Wilcock DM, Morgan D, Gordon MN, Taylor TL, Ridnour LA, Wink DA, Colton CA
    J Neuroinflammation, 2011-09-09;8(0):115.
    Species: Mouse
    Sample Types: Tissue Homogenates
  24. Increased levels of leukocyte-derived MMP-9 in patients with stable angina pectoris.
    Authors: Jonasson S, Lundberg A, Kalvegren H, Bergstrom I, Szymanowski A, Jonasson L
    PLoS ONE, 2011-04-29;6(4):e19340.
    Species: Human
    Sample Types: Serum
  25. 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
  26. Abnormalities in metalloproteinase pathways and IGF-I axis: a link between birth weight, hypertension, and vascular damage in childhood.
    Authors: Sesso R, Franco MC
    Am. J. Hypertens., 2009-11-05;23(1):6-11.
    Species: Human
    Sample Types: Plasma
  27. Salbutamol up-regulates matrix metalloproteinase-9 in the alveolar space in the acute respiratory distress syndrome.
    Authors: O'Kane CM, McKeown SW, Perkins GD, Bassford CR, Gao F, Thickett DR, McAuley DF
    Crit. Care Med., 2009-07-01;37(7):2242-9.
    Species: Human
    Sample Types: BALF
  28. Immunomodulation by chronobiologically-based glucocorticoids treatment for multiple sclerosis relapses.
    Authors: Glass-Marmor L, Paperna T, Galboiz Y, Miller A
    J. Neuroimmunol., 2009-03-28;210(1):124-7.
    Species: Human
    Sample Types: Serum
  29. Imbalance of tissue inhibitors of metalloproteinases (TIMP) - 1 and - 4 serum levels, in patients with inflammatory bowel disease.
    Authors: Kapsoritakis AN, Kapsoritaki AI, Davidi IP, Lotis VD, Manolakis AC, Mylonis PI, Theodoridou AT, Germenis AE, Potamianos SP
    BMC Gastroenterol, 2008-11-26;8(1):55.
    Species: Human
    Sample Types: Serum
  30. MMP expression and abnormal lung permeability are important determinants of outcome in IPF.
    Authors: McKeown S, Richter AG, O'Kane C, O'Kane C, McAuley DF, Thickett DR
    Eur. Respir. J., 2008-10-01;33(1):77-84.
    Species: Human
    Sample Types: BALF
  31. Impact of acute exposure to tobacco smoke on gelatinases in the bronchoalveolar space.
    Authors: Glader P, Eldh B, Bozinovski S, Andelid K, Sjostrand M, Malmhall C, Anderson GP, Riise GC, Qvarfordt I, Linden A
    Eur. Respir. J., 2008-04-16;32(3):644-50.
    Species: Human
    Sample Types: BALF
  32. Is metalloproteinase-7 specific for idiopathic pulmonary fibrosis?
    Authors: Huh JW, Kim DS, Oh YM, Shim TS, Lim CM, Lee SD, Koh Y, Kim WS, Kim WD, Kim KR
    Chest, 2007-12-10;133(5):1101-6.
    Species: Human
    Sample Types: BALF
  33. Actinobacillus actinomycetemcomitans lipopolysaccharide regulates matrix metalloproteinase, tissue inhibitors of matrix metalloproteinase, and plasminogen activator production by human gingival fibroblasts: a potential role in connective tissue destruction.
    Authors: Bodet C, Andrian E, Tanabe S, Grenier D
    J. Cell. Physiol., 2007-07-01;212(1):189-94.
    Species: Human
    Sample Types: Cell Culture Supernates
  34. Matrix metalloproteinase-2 dysregulation in type 1 diabetes.
    Authors: Thrailkill KM, Bunn RC, Moreau CS, Cockrell GE, Simpson PM, Coleman HN, Frindik JP, Kemp SF, Fowlkes JL
    Diabetes Care, 2007-06-11;30(9):2321-6.
    Species: Human
    Sample Types: Plasma
  35. Plasma MMP-2 and MMP-9 and their inhibitors TIMP-1 and TIMP-2 during human orthotopic liver transplantation. The effect of aprotinin and the relation to ischemia/reperfusion injury.
    Authors: Kuyvenhoven JP, Molenaar IQ, Verspaget HW, Veldman MG, Palareti G, Legnani C, Moolenburgh SE, Terpstra OT, Lamers CB, van Hoek B, Porte RJ
    Thromb. Haemost., 2004-03-01;91(3):506-13.
    Species: Human
    Sample Types: Plasma
  36. Matrix metalloproteinases and tissue inhibitors of metalloproteinases in sera and tissue of patients with Dupuytren's disease.
    Authors: Ulrich D, Hrynyschyn K, Pallua N
    Plast. Reconstr. Surg., 2003-10-01;112(5):1279-86.
    Species: Human
    Sample Types: Serum

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Human TIMP-2 Quantikine ELISA Kit
By Anonymous on 05/04/2020
Sample Tested: Urine