Recombinant Human TIMP-1 Protein, CF

R&D Systems | Catalog # 970-TM

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human TIMP-1 Protein (970-TM)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

Q6FGX5

Applications

Inhibition Activity
View all TIMP-1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human TIMP-1 protein
Cys24-Ala207

Purity

>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.

N-terminal Sequence Analysis

Cys24

Predicted Molecular Mass

21 kDa

SDS-PAGE

28-30 kDa, reducing conditions

Activity

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 approximately 2.5 nM as measured under the described conditions. 

Reviewed Applications

Read 4 reviews rated 4.5 using 970-TM in the following applications:

Formulation, Preparation, and Storage

970-TM
Formulation Lyophilized from a 0.2 μm filtered solution in Sodium Acetate and NaCl.
Reconstitution

Reconstitute at 100-500 μg/mL in sterile, deionized water.
Shipping The product is shipped at ambient temperature. 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 28 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

Alternate Names

TIMP1

Entrez Gene IDs

7076 (Human); 21857 (Mouse); 116510 (Rat)

Gene Symbol

TIMP1

UniProt

Q6FGX5

Additional TIMP-1 Products

Product Documents for Recombinant Human TIMP-1 Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human TIMP-1 Protein, CF

For research use only

Citations for Recombinant Human TIMP-1 Protein, CF

Customer Reviews for Recombinant Human TIMP-1 Protein, CF (4)

4.5 out of 5
4 Customer Ratings
5 Stars
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4 Stars
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3 Stars
25%
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1 Stars
0%

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  • Recombinant Human TIMP-1 Protein, CF
    Name: Anonymous
    Application: IP for LC/MS analysis.
    Verified Customer | Posted 07/12/2018
  • Name: Balaji Mahender
    Application: Immunoassay Standard
    Verified Customer | Posted 12/20/2017
  • Recombinant Human TIMP-1 Protein, CF
    Name: Anonymous
    Application: Immunoassay Standard
    Verified Customer | Posted 11/16/2017
    This recombinant protein was used to build an ELISA standard while using MAB970 and BAF970 as the matched pair antibodies.
    Recombinant Human TIMP-1 Protein, CF 970-TM
  • Recombinant Human TIMP-1 Protein, CF
    Name: Anonymous
    Application: Immunoassay Standard
    Verified Customer | Posted 05/25/2017

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Protocols

View specific protocols for Recombinant Human TIMP-1 Protein, CF (970-TM):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35 (w/v), pH 7.5 (TCNB)
  • Recombinant Human TIMP-1 (rhTIMP-1) (Catalog # 970-TM)
  • 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 of APMA at final concentration respectively, in Assay Buffer.
  2. Incubate activation of rhMMP-2 at 37°C for 1 hour.
  3. Prepare a curve of rhTIMP-1 (MW: 20,695 Da). Make the following serial dilutions in Assay Buffer: 2000 nM, 1,000 nM, 500 nM, 300 nM, 200 nM, 150 nM, 100 nM, 20 nM, and 2 nM.
  4. Dilute activated 100 µg/mL rhMMP-2 to 12.8 µg/mL in Assay Buffer.
  5. Mix 25 µL of 12.8 µg/mL rhMMP-2, 16 µL of rhTIMP-1 serial curve dilutions, and 119 µL of Assay Buffer in micro-tubes.
  6. Include two enzyme controls of 25 µL of 12.8 µg/mL rhMMP-2 and 135 mL Assay Buffer in micro-tubes.
  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 5-fold 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 IC50 value of rhTIMP-1 from the curve.
  14. Calculate specific activity for each point 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
  • rhTIMP-1 curve: 20 nM, 10 nM, 5 nM, 3 nM, 2 nM, 1.5 nM, 1.0 nM, 0.2 nM, 0.02 nM, and 0 nM
  • Substrate: 5 µM

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