Recombinant Human MMP-10 Protein, CF

Catalog # Availability Size / Price Qty
910-MP-010
R&D Systems Recombinant Proteins and Enzymes
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Product Details
Citations (4)
FAQs
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Recombinant Human MMP-10 Protein, CF Summary

Product Specifications

Purity
>90%, 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.
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (Catalog # ES002). The specific activity is >190 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human MMP-10 protein
Tyr18-Cys476
Accession #
N-terminal Sequence
Analysis
Tyr18
Structure / Form
Pro and active forms
Predicted Molecular Mass
52 kDa (pro) & 43 kDa (active)
SDS-PAGE
58 kDa and 48 kDa, reducing conditions

Product Datasheets

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910-MP

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.

910-MP

Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl, CaCl2 and Brij-35.
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 opening.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
  • Recombinant Human MMP-10 (rhMMP-10) (Catalog # 910-MP)
  • p-Aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563) 100 mM stock in DMSO
  • Substrate: MCA-Arg-Pro-Lys-Pro-Val-Glu-NVAL-Trp-Arg-Lys(DNP)-NH2 (Catalog # ES002)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: Gemini EM by Molecular Devices) or equivalent
  1. Activate rhMMP-10 at 100 µg/mL in Assay Buffer containing 1 mM APMA.
  2. Incubate at 37 °C for 2 hours.
  3. Dilute activated rhMMP-10 to 2 ng/µL in Assay Buffer.
  4. Dilute Substrate to 20 µM in Assay Buffer.
  5. Load 50 µL of the 2 ng/µL rhMMP-10 in a black well plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate without any rhMMP-10.
  6. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. Calculate specific activity:

     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-10: 0.1 µg
  • Substrate: 10 µM
Reconstitution Calculator

Reconstitution Calculator

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: MMP-10

Matrix metalloproteinases are a family of zinc and calcium dependent endopeptidases with the combined ability to degrade all the components of the extracellular matrix. MMP-10 (stromelysin 2) degrades a broad range of substrates including gelatin, collagen types III, IV and V, fibronectin, aggrecan, and pig cartilage proteoglycan. MMP-10 can activate other MMPs such as MMP-1 and MMP-8. MMP-10 is expressed in keratinocytes, T cells, menstrual endometrium and a few tumor samples. Structurally, MMP-10 may be divided into four distinct domains: a pro-domain which is cleaved upon activation, a catalytic domain containing the zinc binding site; a short linker region, and a carboxyl terminal hemopexin-like domain.

Long Name
Matrix Metalloproteinase 10
Entrez Gene IDs
4319 (Human)
Alternate Names
EC 3.4.24; EC 3.4.24.22; matrix metallopeptidase 10 (stromelysin 2); matrix metalloprotease 10; matrix metalloproteinase 10 (stromelysin 2); Matrix metalloproteinase-10; MMP10; MMP-10; SL-2; STMY2stromelysin-2; Stromelysin 2; transin 2; transin-2

Citations for Recombinant Human MMP-10 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.

4 Citations: Showing 1 - 4
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  1. Functional human iPSC-derived alveolar-like cells cultured in a miniaturized 96?Transwell air-liquid interface model
    Authors: T Bluhmki, S Traub, AK Müller, S Bitzer, E Schruf, MT Bammert, M Leist, F Gantner, JP Garnett, R Heilker
    Scientific Reports, 2021-08-23;11(1):17028.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  2. Matrix metalloproteinase-10 protects against acute kidney injury by augmenting epidermal growth factor receptor signaling
    Authors: C Hu, Y Zuo, Q Ren, X Sun, S Zhou, J Liao, X Hong, J Miao, L Zhou, Y Liu
    Cell Death & Disease, 2021-01-12;12(1):70.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  3. Biodistribution of Nanostructured Lipid Carriers in Mice Atherosclerotic Model
    Authors: L Devel, G Almer, C Cabella, F Beau, M Bernes, P Oliva, F Navarro, R Prassl, H Mangge, I Texier
    Molecules, 2019-09-26;24(19):.
    Species: Human
    Sample Types: Peptide
    Applications: Bioassay
  4. Simple pseudo-dipeptides with a P2' glutamate: a novel inhibitor family of matrix metalloproteases and other metzincins.
    Authors: Devel L, Beau F, Amoura M, Vera L, Cassar-Lajeunesse E, Garcia S, Czarny B, Stura E, Dive V
    J Biol Chem, 2012-06-11;287(32):26647-56.
    Applications: Enzyme Assay

FAQs

  1. Can the enzyme be stored after activation, or do I need to use it immediately after activation?

    • We recommend only activating the amount of enzyme needed for your assay, and recommend activating the enzyme immediately prior to use. Any unactivated enzyme should be stored in aliquots at either the stock concentration at which the enzyme was supplied, or the reconstitution concentration, according to the product datasheet.

  2. If I use this enzyme at a higher concentration, do I need to change the concentration of APMA to activate it?

    • We have only optimized activation conditions for one particular concentration of this MMP enzyme as part of our regular QC testing for enzymatic activity. Activating the enzyme at any different concentration would have to be optimized by the end user.

  3. Does this MMP enzyme need to be activated to work?

    • Yes, this enzyme requires activation prior to use.

  4. What is the ratio of pro- and active forms of this enzyme prior to activation?

    • We do not measure the ratio of pro- to active MMP-10 prior to activation. We measure the activity of each lot after activation to ensure it passes our activity specification.

  5. What is the activity of this enzyme in units/µg?

    • We supply this enzyme as a mass and calculate its activity relative to mass (pmol/min/µg). We have not calibrated this enzyme to an international standard unit, so we are unable to provide a conversion to units/µg.

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Recombinant Human MMP-10 Protein, CF
By Oleg Yerov on 11/06/2020
Application: Enzymatic activity in vitro