Mouse TIMP-1 Quantikine ELISA Kit

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MTM100
Mouse TIMP-1 ELISA Standard Curve
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Product Details
Procedure
Citations (17)
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Mouse TIMP-1 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 (17 uL), Serum (17 uL), EDTA Plasma (17 uL)
Sensitivity
3.5 pg/mL
Assay Range
37.5 - 2,400 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
Specificity
Natural and recombinant mouse TIMP-1. This kit detects approximately 80% of the recombinant mouse TIMP-1 complexed with the pro-form of recombinant mouse MMP-9 and approximately 60% when complexed with active recombinant mouse MMP-9.
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 Mouse TIMP-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse TIMP-1 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse TIMP-1 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse TIMP-1 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative levels of natural mouse TIMP-1.

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, EDTA Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 91.8 282 721 89.7 293 721
Standard Deviation 9.4 12.6 23 6.5 22.1 41.4
CV% 10.2 4.5 3.2 7.2 7.5 5.7

Recovery

The recovery of mouse TIMP-1 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=6) 102 92-112
EDTA Plasma (n=4) 107 92-119
Serum (n=6) 101 84-117

Linearity

To assess the linearity of the assay, samples containing high concentrations of mouse TIMP-1 in each matrix were diluted with Calibrator Diluent and assayed.
Mouse TIMP-1 ELISA Linearity

Data Examples

Mouse TIMP-1 ELISA Standard Curve

Product Datasheets

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-1

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 1
Entrez Gene IDs:
7076 (Human); 21857 (Mouse); 116510 (Rat)
Alternate Names:
CLGI; Collagenase inhibitor; collagenase inhibitor); EPATIMP-1; EPO; erythroid potentiating activity; Erythroid-potentiating activity; Fibroblast collagenase inhibitor; FLJ90373; HCI; metalloproteinase inhibitor 1; TIMP metallopeptidase inhibitor 1; TIMP1; TIMP-1; TIMPtissue inhibitor of metalloproteinase 1 (erythroid potentiating activity; Tissue inhibitor of metalloproteinases 1
⚠ 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. 50 µL Assay Diluent
  4.   Add 50 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.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 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 Mouse TIMP-1 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.

17 Citations: Showing 1 - 10
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  1. SILAC Analysis Reveals Increased Secretion of Hemostasis-Related Factors by Senescent Cells
    Authors: CD Wiley, S Liu, C Limbad, AM Zawadzka, J Beck, M Demaria, R Artwood, F Alimirah, JA Lopez-Domi, C Kuehnemann, SR Danielson, N Basisty, HG Kasler, TR Oron, PY Desprez, SD Mooney, BW Gibson, B Schilling, J Campisi, P Kapahi
    Cell Rep, 2019;28(13):3329-3337.e5.
    Species: Mouse
    Sample Types: Platelet-Poor Plasma
  2. Gemcitabine-induced TIMP1 attenuates therapy response and promotes tumor growth and liver metastasis in pancreatic cancer
    Authors: Z D'Costa, K Jones, A Azad, R van Stipho, SY Lim, AL Gomes, P Kinchesh, SC Smart, WG McKenna, FM Buffa, OJ Sansom, RJ Muschel, E O'Neill, E Fokas
    Cancer Res., 2017;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  3. Monocyte chemotactic protein-1 deficiency reduces spontaneous metastasis of Lewis lung carcinoma in mice fed a high-fat diet
    Oncotarget, 2016;7(17):24792-9.
    Species: Mouse
    Sample Types: Tissue Homogenates
  4. Effects of a high-fat diet on spontaneous metastasis of Lewis lung carcinoma in plasminogen activator inhibitor-1 deficient and wild-type mice.
    Authors: Yan L, DeMars L
    PLoS ONE, 2014;9(10):e110869.
    Species: Mouse
    Sample Types: Plasma
  5. CXCL6 antibody neutralization prevents lung inflammation and fibrosis in mice in the bleomycin model.
    Authors: Besnard A, Struyf S, Guabiraba R, Fauconnier L, Rouxel N, Proost P, Uyttenhove C, Van Snick J, Couillin I, Ryffel B
    J Leukoc Biol, 2013;94(6):1317-23.
    Species: Mouse
    Sample Types: Tissue Homogenates
  6. Role of c-Jun N-terminal protein kinase 1/2 (JNK1/2) in macrophage-mediated MMP-9 production in response to Moraxella catarrhalis lipooligosaccharide (LOS).
    Authors: Hassan F, Ren D, Zhang W
    PLoS ONE, 2012;7(5):e37912.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  7. The effects of low levels of dystrophin on mouse muscle function and pathology.
    Authors: van Putten M, Hulsker M, Nadarajah VD, van Heiningen SH, van Huizen E, van Iterson M, Admiraal P, Messemaker T, den Dunnen JT, 't Hoen PA, Aartsma-Rus A
    PLoS ONE, 2012;7(2):e31937.
    Species: Mouse
    Sample Types: Serum
  8. 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;8(0):115.
    Species: Mouse
    Sample Types: Tissue Homogenates
  9. Tissue inhibitor of matrix metalloproteinase 1 (TIMP1) controls adipogenesis in obesity in mice and in humans.
    Authors: Meissburger B, Stachorski L, Roder E, Rudofsky G, Wolfrum C
    Diabetologia, 2011;54(6):1468-79.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. Adenosine and osteopontin contribute to the development of chronic obstructive pulmonary disease.
    Authors: Schneider DJ, Lindsay JC, Zhou Y, Molina JG, Blackburn MR
    FASEB J., 2010;24(1):70-80.
    Species: Mouse
    Sample Types: BALF
  11. Integrated proteomic analysis of human cancer cells and plasma from tumor bearing mice for ovarian cancer biomarker discovery.
    Authors: Pitteri SJ, JeBailey L, Faca VM, Thorpe JD, Silva MA, Ireton RC, Horton MB, Wang H, Pruitt LC, Zhang Q, Cheng KH, Urban N, Hanash SM, Dinulescu DM
    PLoS ONE, 2009;4(11):e7916.
    Species: Mouse
    Sample Types: Plasma
  12. Retinoic acid decreases adherence of murine myeloid dendritic cells and increases production of matrix metalloproteinase-9.
    Authors: Lackey DE, Ashley SL, Davis AL, Hoag KA
    J. Nutr., 2008;138(8):1512-9.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  13. A3 adenosine receptor signaling influences pulmonary inflammation and fibrosis.
    Authors: Morschl E, Molina JG, Volmer JB, Mohsenin A, Pero RS, Hong JS, Kheradmand F, Lee JJ, Blackburn MR
    Am. J. Respir. Cell Mol. Biol., 2008;39(6):697-705.
    Species: Mouse
    Sample Types: BALF
  14. Atheroprotective role of interleukin-6 in diet- and/or pathogen-associated atherosclerosis using an ApoE heterozygote murine model.
    Authors: Madan M, Bishayi B, Hoge M, Amar S
    Atherosclerosis, 2007;197(2):504-14.
    Species: Mouse
    Sample Types: Serum
  15. Amelioration of coxsackievirus B3-mediated myocarditis by inhibition of tissue inhibitors of matrix metalloproteinase-1.
    Authors: Crocker SJ, Frausto RF, Whitmire JK, Benning N, Milner R, Whitton JL
    Am. J. Pathol., 2007;171(6):1762-73.
    Species: Mouse
    Sample Types: Tissue Homogenates
  16. TIMP-1 gene deficiency increases tumour cell sensitivity to chemotherapy-induced apoptosis.
    Authors: Davidsen ML, Wurtz SØ, Romer MU, Sorensen NM, Johansen SK, Christensen IJ, Larsen JK, Offenberg H, Brunner N, Lademann U
    Br. J. Cancer, 2006;95(8):1114-20.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  17. Effects of bicyclol on dimethylnitrosamine-induced liver fibrosis in mice and its mechanism of action.
    Authors: Hu QW, Liu GT
    Life Sci., 2006;79(6):606-12.
    Species: Mouse
    Sample Types: Serum

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