Recombinant Rat TIMP-1 Protein, CF

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R&D Systems Recombinant Proteins and Enzymes
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Recombinant Rat TIMP-1 Protein, CF Summary

Product Specifications

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
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 <3.0 nM, as measured under the described conditions.
Mouse myeloma cell line, NS0-derived rat TIMP-1 protein
Accession #
N-terminal Sequence
Predicted Molecular Mass
21.5 kDa
32-34 kDa, reducing conditions

Product Datasheets

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Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.


Formulation Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.
Reconstitution Reconstitute at 500 μg/mL in sterile, deionized water.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Assay Procedure

  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05 % (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Rat TIMP-1 (rrTIMP-1) (Catalog # 580-RT)
  • 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. Activate rhMMP-2 at 100 µg/mL with 1 mM APMA in Assay Buffer.
  2. Incubate at 37 ºC for 1 hour to activate rhMMP-2.
  3. Dilute activated rhMMP-2 to 12.5 µg/mL in Assay Buffer.
  4. Prepare a curve of rrTIMP-1 (MW: 21,500 Da) in Assay Buffer. Make the following serial dilutions: 5000, 2000, 1000, 500, 300, 200, 150, 100, 20, and 2 nM.
  5. Mix 25.6 µL of 12.5 µg/mL rhMMP-2, 16 µL of rrTIMP-1 serial curve dilutions, and 118.4 µL of Assay Buffer in microtubes. Include two enzyme controls of 25.6 µL of 12.5 µg/mL rhMMP-2 and 134.4 µL Assay Buffer in microtubes.
  6. Incubate reaction mixtures at 37 °C for 2 hours.
  7. Dilute incubated reaction mixtures 5 fold in Assay Buffer.
  8. Dilute Substrate to 10 µM in Assay Buffer.
  9. In a plate load 50 µL of the diluted incubated reaction mixtures to wells, and start the reaction by adding 50 µL of 10 µM Substrate.
  10. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  11. Determine the 50% inhibition concentration (IC50) for rrTIMP-1 by plotting RFU/min (or specific activity) vs. concentration with 4‑PL fitting.
  12. The specific activity for rhMMP-2 at each point may be determined using the following formula (if needed):


     Specific Activity (pmoles/min/µg) =

Adjusted Vmax* (RFU/min) x Conversion Factor** (pmole/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
  • rrTIMP-1 curve: 50, 20, 10, 5, 3, 2, 1.5, 1, 0.2, 0.02, and 0 nM
  • Substrate: 5 µM
Reconstitution Calculator

Reconstitution Calculator

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Background: TIMP-1

Tissue inhibitors of metalloproteinases or TIMPs are a family of proteins that regulate the activation and proteolytic activity of the zinc enzymes known as matrix metalloproteinases (MMPs). There are four members of the family, TIMP-1, TIMP-2, TIMP-3 and TIMP-4. TIMP-1 is a glycoprotein with a molecular mass of 32‑34 kDa produced by a wide range of cell types. TIMP-1 inhibits active MMP-mediated proteolysis by forming an N-terminal, non-covalent binary complex with the MMP active site. TIMP-1 also associates C-terminally with pro-MMP-9 in a complex which may play a role in regulating activation. Independent of MMPs, TIMP-1 has been shown to have a role in tissue homeostasis.

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

Citations for Recombinant Rat TIMP-1 Protein, CF

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.

5 Citations: Showing 1 - 5
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  1. Ectodomain shedding of Limbic System-Associated Membrane Protein (LSAMP) by ADAM Metallopeptidases promotes neurite outgrowth in DRG neurons
    Authors: RL Sanz, GB Ferraro, MP Girouard, AE Fournier
    Sci Rep, 2017-08-11;7(1):7961.
    Species: Rat
    Sample Types: Whole Cells
    Applications: Bioassay
  2. Tissue Inhibitor Of Matrix Metalloproteinase-1 Is Required for High-Fat Diet-Induced Glucose Intolerance and Hepatic Steatosis in Mice.
    Authors: Fjaere E, Andersen C, Myrmel L, Petersen R, Hansen J, Tastesen H, Mandrup-Poulsen T, Brunner N, Kristiansen K, Madsen L, Romer M
    PLoS ONE, 2015-07-13;10(7):e0132910.
    Species: Rat
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  3. Adiponectin reduces hepatic stellate cell migration by promoting tissue inhibitor of metalloproteinase-1 (TIMP-1) secretion.
    Authors: Ramezani-Moghadam M, Wang J, Ho V, Iseli T, Alzahrani B, Xu A, Van der Poorten D, Qiao L, George J, Hebbard L
    J Biol Chem, 2015-01-09;290(9):5533-42.
    Species: Rat
    Sample Types: Recombinant Protein, Whole Cells
    Applications: Bioassay, Western Blot
  4. IgLON cell adhesion molecules are shed from the cell surface of cortical neurons to promote neuronal growth.
    Authors: Sanz R, Ferraro G, Fournier A
    J Biol Chem, 2014-12-23;290(7):4330-42.
    Species: Rat
    Sample Types: Whole Cells
    Applications: Bioassay
  5. Activated hepatic stellate cells are dependent on self-collagen, cleaved by membrane type 1 matrix metalloproteinase for their growth.
    Authors: Birukawa N, Murase K, Sato Y, Kosaka A, Yoneda A, Nishita H, Fujita R, Nishimura M, Ninomiya T, Kajiwara K, Miyazaki M, Nakashima Y, Ota S, Murakami Y, Tanaka Y, Minomi K, Tamura Y, Niitsu Y
    J Biol Chem, 2014-05-27;289(29):20209-21.
    Species: Rat
    Sample Types: Whole Cells
    Applications: Bioassay


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