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Recombinant Mouse TIMP-1 Protein, CF

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Summary
Reactivity MuSpecies Glossary
Applications Inhibition Activity
Format
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Recombinant Mouse TIMP-1 Protein, CF Summary

Details of Functionality
Measured by its ability to inhibit human MMP-2 cleavage of a fluorogenic peptide substrate Mca-PLGL-Dpa-AR-NH2 (Catalog # ES001). The IC50 value is <3.5 nM, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived mouse TIMP-1 protein
Cys25-Arg205
Accession #
N-terminal Sequence
Cys25
Protein/Peptide Type
Recombinant Enzymes
Gene
Timp1
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Inhibition Activity
Theoretical MW
20 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
32 kDa, reducing conditions
Publications
Read Publications using
980-MT in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Buffer
Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Reconstitution Instructions
Reconstitute at 100 μg/mL in sterile 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, and 0.05% Brij-35, pH 7.5.
Assay Procedure
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35 (w/v), pH 7.5 (TCNB)
  • Recombinant Mouse TIMP-1 (rmTIMP-1) (Catalog # 980-MT)
  • Recombinant Human MMP‑2 (rhMMP‑2) (Catalog # 902-MP)
  • p-aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563), 100 mM stock in DMSO
  • Substrate: MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES001) , 2 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhMMP-2 to 100 µg/mL in Assay Buffer containing 1 mM APMA.
  2. Incubate at 37 °C for 1 hour (activation).
  3. Prepare a curve of rmTIMP-1 (MW: 20,200 Da). Make the following serial dilutions in Assay Buffer:  5000, 2000, 1000, 500, 300, 200, 150, 100, 20, and 2 nM.
  4. Dilute activated rhMMP-2 to 12.5 µg/mL in Assay Buffer.
  5. Mix 25.6 µL of diluted rhMMP-2, 16 µL of rmTIMP-1 serial curve dilutions, and 118.4 µL of Assay Buffer.
  6. Include two enzyme controls of 25.6 µL of diluted rhMMP-2 and 134.4 μL Assay Buffer.
  7. Incubate reaction mixtures at 37 °C for 2 hours.
  8. Dilute incubated reaction mixtures by a 5-fold dilution in Assay Buffer.
  9. Dilute Substrate to 10 µM in Assay Buffer.
  10. In a plate load 50 µL of the diluted incubated reaction mixtures to wells.
  11. Start the reaction by adding 50 µL of 10 µM Substrate to wells.
  12. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively in kinetic mode for 5 minutes.
  13. Derive the 50% inhibiting concentration (IC50) for rmTIMP-1 by plotting RFU/min (or specific activity) vs. concentration with 4-PL fitting.
  14. The specific activity for rhMMP-2 at each point may be determined using the following formula (if needed):

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

Per Well:
  • rhMMP-2:  0.02 µg
  • rmTIMP-1:  50, 20, 10, 5, 3, 2, 1.5, 1, 0.2, and 0.02 nM
  • Substrate: 5 µM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse TIMP-1 Protein, CF

  • 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

Background

Tissue inhibitors of metalloproteinases or TIMPs are a family of homologous proteins that regulate the activity of matrix metalloproteinases (MMPs) (1, 2). There are four known members of the family, TIMP‑1, TIMP-2, TIMP-3 and TIMP-4 that have been found to exhibit multiple functions, including inhibition of active MMPs, proMMP activation, cell growth promotion, matrix binding, inhibition of angiogenesis and the induction of apoptosis. Structurally, TIMPs have two domains, an N‑terminal domain and a C‑terminal domain. Each domain consists of three disulfide-bonded loops. TIMP-1 is a glycoprotein produced by a wide range of cell types. Through its N‑terminal domain, TIMP-1 inhibits active MMPs by forming a non-covalent binary complex with the MMP active site. The C-terminal domain of TIMP-1 interacts with the C-terminal domain of proMMP-9, which may play a role in regulating proMMP-9 activation.

  1. Murphy, G. and F. Willenbrock (1995) Methods Enzymol. 248:496.
  2. Brew, K. et al. (2000) Biochim. Biophys. Acta 1477:267.

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Recombinant Mouse
TIMP-1 Protein, CF
980-MT
Recombinant Mouse TIMP-1 Protein, CF
Species: Mu
Applications: Inhibition Activity

Publications for TIMP-1 (980-MT)(14)

We have publications tested in 3 confirmed species: Mouse, Hamster, Transgenic Mouse.

We have publications tested in 3 applications: Bioassay, In Vivo, In vivo assay.


Filter By Application
Bioassay
(12)
In Vivo
(1)
In vivo assay
(1)
All Applications
Filter By Species
Mouse
(12)
Hamster
(1)
Transgenic Mouse
(4)
All Species
Showing Publications 1 - 10 of 14. Show All 14 Publications.
Publications using 980-MT Applications Species
Priego, N;de Pablos-Aragoneses, A;Perea-García, M;Pieri, V;Hernandez-Oliver, C;Alvaro-Espinosa, L;Rojas, A;Sanchez, O;Steindl, A;Caleiras, E;Garcia, F;Garcia-Martin, S;Grana-Castro, O;Garcia-Mulero, S;Serrano, D;Velasco-Beltran, P;Jimenez-Lasheras, B;Egia-Mendikute, L;Rupp, L;Stammberger, A;Meinhardt, M;Chaachou-Charradi, A;Martínez-Saez, E;Bertero, L;Cassoni, P;Mangherini, L;Pellerino, A;Ruda, R;Soffietti, R;Al-Shahrour, F;Saftig, P;Sanz-Pamplona, R;Schmitz, M;Crocker, SJ;Calvo, A;Palazon, A;Group, R;Valiente, M; TIMP1 mediates astrocyte-dependent local immunosuppression in brain metastasis acting on infiltrating CD8+ T cells Cancer discovery 2024-10-02 [PMID: 39354883] (Bioassay, Mouse) Bioassay Mouse
Nakamura, K;Tsukasaki, M;Tsunematsu, T;Yan, M;Ando, Y;Huynh, NC;Hashimoto, K;Gou, Q;Muro, R;Itabashi, A;Iguchi, T;Okamoto, K;Nakamura, T;Nakano, K;Okamura, T;Ueno, T;Ito, K;Ishimaru, N;Hoshi, K;Takayanagi, H; The periosteum provides a stromal defence against cancer invasion into the bone Nature 2024-08-21 [PMID: 39169177] (In vivo assay, Mouse, Transgenic Mouse) In vivo assay Mouse, Transgenic Mouse
Sutter, PA;Willis, CM;Menoret, A;Nicaise, AM;Sacino, A;Sikkema, AH;Jellison, ER;Win, KK;Han, DK;Church, W;Baron, W;Vella, AT;Crocker, SJ; Astrocytic TIMP-1 regulates production of Anastellin, an inhibitor of oligodendrocyte differentiation and FTY720 responses Proceedings of the National Academy of Sciences of the United States of America 2024-01-30 [PMID: 38266047] (Bioassay, Mouse, Transgenic Mouse) Bioassay Mouse, Transgenic Mouse
Geou-Yarh Liou, Alicia K. Fleming Martinez, Heike R. Döppler, Ligia I. Bastea, Peter Storz Inflammatory and alternatively activated macrophages independently induce metaplasia but cooperatively drive pancreatic precancerous lesion growth iScience 2023-06-16 [PMID: 37250781] (Bioassay, Mouse) Bioassay Mouse
PA Sutter, CM Willis, A Menoret, AM Nicaise, A Sacino, AH Sikkema, E Jellison, KK Win, DK Han, W Church, W Baron, AT Vella, SJ Crocker Astrocytic TIMP-1 regulates production of Anastellin, a novel inhibitor of oligodendrocyte differentiation and FTY720 responses bioRxiv : the preprint server for biology, 2023-02-18;0(0):. 2023-02-18 [PMID: 36824834] (Bioassay, Transgenic Mouse) Bioassay Transgenic Mouse
F Girardi, A Taleb, M Ebrahimi, A Datye, DG Gamage, C Peccate, L Giordani, DP Millay, PM Gilbert, B Cadot, F Le Grand TGF&amp;beta signaling curbs cell fusion and muscle regeneration Nature Communications, 2021-02-02;12(1):750. 2021-02-02 [PMID: 33531466] (Bioassay, Mouse) Bioassay Mouse
X Wang, J Rojas-Quin, J Wilder, Y Tesfaigzi, D Zhang, CA Owen Tissue Inhibitor of Metalloproteinase-1 Promotes Polymorphonuclear Neutrophil (PMN) Pericellular Proteolysis by Anchoring Matrix Metalloproteinase-8 and -9 to PMN Surfaces J. Immunol., 2019-04-24;0(0):. 2019-04-24 [PMID: 31019060] (Bioassay, Transgenic Mouse) Bioassay Transgenic Mouse
LJ Martin, SB Smith, A Khoutorsky, CA Magnussen, A Samoshkin, RE Sorge, C Cho, N Yosefpour, S Sivaselvac, S Tohyama, T Cole, TM Khuong, E Mir, DG Gibson, JS Wieskopf, SG Sotocinal, JS Austin, CB Meloto, JH Gitt, C Gkogkas, N Sonenberg, JD Greenspan, RB Fillingim, R Ohrbach, GD Slade, C Knott, R Dubner, AG Nackley, A Ribeiro-da, GG Neely, W Maixner, DV Zaykin, JS Mogil, L Diatchenko Epiregulin and EGFR interactions are involved in pain processing J. Clin. Invest., 2017-08-07;0(0):. 2017-08-07 [PMID: 28783046] (In Vivo, Mouse) In Vivo Mouse
F Strålberg, A Kassem, F Kasprzykow, M Abrahamson, A Grubb, C Lindholm, UH Lerner Inhibition of lipopolysaccharide-induced osteoclast formation and bone resorption in vitro and in vivo by cysteine proteinase inhibitors J. Leukoc. Biol, 2017-02-14;0(0):. 2017-02-14 [PMID: 28196851] (Bioassay, Mouse) Bioassay Mouse
Thevenard J, Verzeaux L, Devy J, Etique N, Jeanne A, Schneider C, Hachet C, Ferracci G, David M, Martiny L, Charpentier E, Khrestchatisky M, Rivera S, Dedieu S, Emonard H Low-density lipoprotein receptor-related protein-1 mediates endocytic clearance of tissue inhibitor of metalloproteinases-1 and promotes its cytokine-like activities. PLoS ONE, 2014-07-30;9(7):e103839. 2014-07-30 [PMID: 25075518] (Bioassay, Hamster, Mouse) Bioassay Hamster, Mouse
Show All 14 Publications.

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Bioinformatics

Gene Symbol Timp1
Uniprot