RayBio® Human TIMP-2 ELISA Kit for cell culture supernatants, plasma, and serum samples.

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Catalog #:
ELH-TIMP2
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Product Description

Specifications

Size 1 Plate Kit, 2 Plate Kit, 5 Plate Kit
Species Human
Accession Number P16035
Gene Id 7077
Gene Symbols TIMP2
Protein Name / Synonyms Metalloproteinase inhibitor 2 (CSC-21K) (Tissue inhibitor of metalloproteinases 2) (TIMP-2)
Quantitative/Semi-Quantitative Quantitative
Specificity This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, G-CSF, GM-CSF, IFN-gamma, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, PDGF, SCF, TNF-alpha, TNF-beta, TPO, VEGF.
Compatible Sample Types Cell Culture Supernatants, Plasma, Serum
Solid Support 96-well Microplate
Method Of Detection Colorimetric
Design Principle Sandwich-based
Sensitivity 2 pg/ml

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Detection Range 2 pg/ml - 1200 pg/ml
Estimated Lead Time 1-2 business days
Shipping Type Blue ice
Storage ≤-20°C

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Target Background

Tissue inhibitor of metalloproteinase-2 (TIMP2) is a 21 kDa protein mainly expressed in melanoma and renal tubular cells. TIMP2 inhibits matrix metalloproteinases by binding to their catalytic zinc cofactor. The expression of TIMP2 can be induced by cytokines, chemokines and factors involved in proliferation and differentiation. TIMP2 is involved in G1 cell cycle arrest during the early phase of cell injury. Insulin growth factor binding protein-7 (IGFBP7) is also a marker for cell cycle arrest. TIMP2 together with IGFBP7 were identified as biomarkers for acute kidney injury.

Product Features

Additional Notes
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data

Standard Curves

Spiking & Recovery Results

Linearity Results

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Other Materials Required
  • Distilled or deionized water
  • Precision pipettes to deliver 2 µl to 1 µl volumes
  • Adjustable 1-25 µl pipettes for reagent preparation
  • 100 µl and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Protocol Outline
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 µl of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4°C.
  4. Add 100 µl of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.

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Storage/Stability

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Rottner M, Kunzelmann C, Mergey M, Freyssinet JM, Martinez MC. Exaggerated apoptosis and NF-kappa-B activation in pancreatic and tracheal cystic fibrosis cells. FASEB J. 2007;21:2939-2948.
Species:
Human
Sample Type:
Conditioned Media (Pancreatic cancer (PANG-1 line) cell-cultured medium)
Hampel U, Klonisch T, Sel S, et al. Insulin-like Factor 3 Promotes Wound Healing at the Ocular Surface. Endocrinology. 2013;154(6):2034-2045.
Species:
Human
Sample Type:
Conditioned Media (sebaceous cell line, conjunctival epithelial cell line, corneal epithelial cell line)
Malard V, Berenguer F, Prat O, Ruat S, Steinmetz G, Quemeneur E. Global gene expression profiling in human lung cells exposed to cobalt. BMC Genomics. 2007;8:147. doi:10.1186/1271-2164-8-147.
Species:
Human
Sample Type:
Conditioned Media (Human type II epithelial (A549) cell-Cell-conditioned media)
Khadjavi A., Magnetto C., Panariti A., et al. Chitosan-shelled oxygen-loaded nanodroplets abrogate hypoxia dysregulation of human keratinocyte gelatinases and inhibitors: New insights for chronic wound healing. Toxicol Appl Pharmacol. 2015 Aug 1;286(3):198-206. doi: 10.1016/j.taap.2015.04.015
Species:
Human
Sample Type:
Conditioned Media (HaCaT cells keratinocytes)
Basilico N., Magnetto C., D'Alessandro D., et al. Dextran-shelled oxygen-loaded nanodroplets reestablish a normoxia-like pro-angiogenic phenotype and behavior in hypoxic human dermal microvascular endothelium. Toxicol Appl Pharmacol. 2015 Nov 1;288(3):330-8. doi: 10.1016/j.taap.2015.08.005.
Species:
Human
Sample Type:
Conditioned Media (Human dermal endothelial cells (HMEC-1))
Ciaffi L., Cavassini M., Genne D., et al. Switch to etravirine for HIV-positive patients receiving statin treatment: a prospective study. Eur J Clin Invest. 2015 Jul;45(7):720-30. doi: 10.1111/eci.12464.
Species:
Human
Sample Type:
Serum (etravirine treated HIV patients)
Gulino G., Magnetto C., Khadjavi A., et al. Oxygen-Loaded Nanodroplets Effectively Abrogate Hypoxia Dysregulating Effects on Secretion of MMP-9 and TIMP-1 by Human Monocytes. Mediators Inflamm. 2015;2015:964838. doi: 10.1155/2015/964838.
Species:
Human
Sample Type:
Conditioned Media (Human Adherent Monocytes)
Chapman CL, Johnson BD, Vargas NT, Hostler D, Parker MD, Schlader ZJ. Both hyperthermia and dehydration during physical work in the heat contribute to the risk of acute kidney injury. J Appl Physiol (1985). 2020 Apr 1;128(4):715-728. doi: 10.1152/japplphysiol.00787.2019. Epub 2020 Feb 20. PMID: 32078468; PMCID: PMC7191500.
Species:
Human
Sample Type:
Urine
Cai D, Xu Y, Ding R, Qiu K, Zhang R, Wang H, Huang L, Xie X, Yan H, Deng Y, Lin X, Shao J, Luo X, Duan C. Extensive serum biomarker analysis in patients with non-small-cell lung carcinoma. Cytokine. 2020 Feb;126:154868. doi: 10.1016/j.cyto.2019.154868. Epub 2019 Oct 16. PMID: 31629110.
Species:
Human
Sample Type:
Serum
Koob T., et al. Cytokines in single layer amnion allografts compared to multilayeramnion/chorion allografts for wound healing. Journal of Biomedical Materials Research Part B: Applied Biomaterials. Aug 2014, epub ahead of print. DOI: 10.1002/jbm.b.33265
Species:
Human
Sample Type:
Tissue Lysate (Amnion and Chorion Samples)
  1. TIMP-2
    Rating
    100%
    We ran 3-D cell culture supernatants were run with the kit for the first time as feasibility. We had two different media formulations as background against the culture conditions over a month long culture period.

    Review by

    from Miromatrix,

    on

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