Recombinant Human MMP-1 Protein, CF

Catalog # Availability Size / Price Qty
901-MP-010
Product Details
Citations (15)
FAQs
Supplemental Products
Reviews (1)

Recombinant Human MMP-1 Protein, CF Summary

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.
Activity
Measured by its ability to cleave a fluorogenic peptide substrate Mca-KPLGL-Dpa-AR-NH2 (Catalog # ES010). The specific activity is > 400 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human MMP-1 protein
Phe20-Asn469
Accession #
N-terminal Sequence
Analysis
Phe20
Structure / Form
Pro form
Predicted Molecular Mass
52 kDa
SDS-PAGE
52-55 kDa, reducing conditions

Product Datasheets

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.

901-MP

Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl, CaCl2 and Brij-35.
Shipping The product is shipped with dry ice or equivalent. 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-1 (rhMMP-1) (Catalog # 901-MP)
  • p-aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563), 100 mM stock in DMSO
  • Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (R&D Systems, Catalog # ES010)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhMMP-1 to 50 µg/mL in Assay Buffer.
  2. Activate 50 µg/mL rhMMP-1 by adding APMA to a final concentration of 1 mM.
  3. Incubate at 37 °C for 2 hours.
  4. Dilute activated rhMMP-1 to 1 ng/µL in Assay Buffer.
  5. Dilute Substrate to 20 µM in Assay Buffer.
  6. Load into a black well plate 50 µL of 1 ng/µL rhMMP-1 and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer, 50 µL Substrate, and no rhMMP-1.
  7. Read at excitiation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes.
  8. 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-1: 0.050 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-1

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-1 (interstitial collagenase), can degrade a broad range of substrates including types I, II, III, VII, VIII, and X collagens as well as casein, gelatin, alpha ‑1 antitrypsin, myelin basic protein, L-Selectin, pro-TNF, IL-1 beta, IGF-BP3, IGF-BP5, pro MMP-2 and pro MMP-9. A significant role of MMP-1 is the degradation of fibrillar collagens in extracellular matrix remodeling, characterized by the cleavage of the interstitial collagen triple helix into ¾, ¼ fragments. However, as the list of substrates above illustrates, the role of MMP-1 is more diverse than originally envisaged, and may involve enzyme cascades, cytokine regulation and cell surface molecule modulation. MMP-1 is expressed by fibroblasts, keratinocytes, endothelial cells, monocytes and macrophages. Structurally, MMP-1 may be divided into several distinct domains; a pro-domain which is cleaved upon activation; a catalytic domain containing the zinc binding site; a short hinge region and a carboxyl terminal (hemopexin-like) domain.

References
  1. Cawston, T.E. (2004) in Interstitial Collagenase. Barrett, A.J. et al. (eds): Handbook of Proteolytic Enzymes, San Diego: Academic Press, p. 472.
Long Name
Matrix Metalloproteinase 1
Entrez Gene IDs
4312 (Human); 83995 (Mouse)
Alternate Names
CLGmatrix metalloprotease 1; CLGN; EC 3.4.24; EC 3.4.24.7; Fibroblast collagenase; interstitial collagenase; matrix metallopeptidase 1 (interstitial collagenase); matrix metalloproteinase 1 (interstitial collagenase); Matrix metalloproteinase-1; MMP1; MMP-1

Citations for Recombinant Human MMP-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.

15 Citations: Showing 1 - 10
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  1. Efficient protease based purification of recombinant matrix metalloprotease-1 in E. coli
    Authors: L Kumar, W Colomb, J Czerski, CR Cox, SK Sarkar
    Protein Expr. Purif., 2018;0(0):.
    Applications: Enzyme Assay
  2. Nidogen-1 Degraded by Cathepsin S can be Quantified in Serum and is Associated with Non-Small Cell Lung Cancer
    Authors: N Willumsen, CL Bager, DJ Leeming, AC Bay-Jensen, MA Karsdal
    Neoplasia, 2017;19(4):271-278.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  3. MMP-1 Activation Contributes to Airway Smooth Muscle Growth and Asthma Severity
    Authors: Shams-Un-Nisa Naveed
    Am. J. Respir. Crit. Care Med, 2016;0(0):.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  4. Transformed MDCK cells secrete elevated MMP1 that generates LAMA5 fragments promoting endothelial cell angiogenesis
    Authors: Shashi K Gopal
    Sci Rep, 2016;6(0):28321.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  5. TLR3 activation augments matrix metalloproteinase production through reactive nitrogen species generation in human lung fibroblasts.
    Authors: Ichikawa T, Sugiura H, Koarai A, Minakata Y, Kikuchi T, Morishita Y, Oka A, Kanai K, Kawabata H, Hiramatsu M, Akamatsu K, Hirano T, Nakanishi M, Matsunaga K, Yamamoto N, Ichinose M
    J Immunol, 2014;192(11):4977-88.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: Bioassay
  6. Fibulin-3, -4, and -5 are highly susceptible to proteolysis, interact with cells and heparin, and form multimers.
    Authors: Djokic J, Fagotto-Kaufmann C, Bartels R, Nelea V, Reinhardt D
    J Biol Chem, 2013;288(31):22821-35.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  7. Resistance of corneal RFUVA-cross-linked collagens and small leucine-rich proteoglycans to degradation by matrix metalloproteinases.
    Authors: Zhang Y, Mao X, Schwend T, Littlechild S, Conrad G
    Invest Ophthalmol Vis Sci, 2013;54(2):1014-25.
    Species: Bovine
    Sample Types: Protein
    Applications: Enzyme Assay
  8. 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;287(32):26647-56.
    Species: N/A
    Sample Types: Fluorogenic Peptide Substrate
    Applications: Enzyme Assay
  9. Pre-analytical effects of blood sampling and handling in quantitative immunoassays for rheumatoid arthritis.
    Authors: Zhao X, Qureshi F, Eastman PS, Manning WC, Alexander C, Robinson WH, Hesterberg LK
    J. Immunol. Methods, 2012;378(1):72-80.
    Species: N/A
    Sample Types: N/A
    Applications: ELISA (Standard)
  10. Development and validation of sandwich ELISA microarrays with minimal assay interference.
    Authors: Gonzalez RM, Seurynck-Servoss SL, Crowley SA
    J. Proteome Res., 2008;7(6):2406-14.
    Species: N/A
    Sample Types: N/A
    Applications: ELISA (Standard)
  11. Interleukin-1beta induced activation of nuclear factor-kappab can be inhibited by novel pharmacological agents in osteoarthritis.
    Authors: Lauder SN, Carty SM, Carpenter CE
    Rheumatology (Oxford), 2007;46(5):752-8.
    Species: N/A
    Sample Types: N/A
    Applications: ELISA (Standard)
  12. Cleavage of myelin associated glycoprotein by matrix metalloproteinases.
    Authors: Milward E, Kim KJ, Szklarczyk A, Nguyen T, Melli G, Nayak M, Deshpande D, Fitzsimmons C, Hoke A, Kerr D, Griffin JW, Calabresi PA, Conant K
    J. Neuroimmunol., 2007;193(1):140-8.
    Species: Rat
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  13. Targeting ADAM-mediated ligand cleavage to inhibit HER3 and EGFR pathways in non-small cell lung cancer.
    Authors: Zhou BB, Peyton M, He B, Liu C, Girard L, Caudler E, Lo Y, Baribaud F, Mikami I, Reguart N, Yang G, Li Y, Yao W, Vaddi K, Gazdar AF, Friedman SM, Jablons DM, Newton RC, Fridman JS, Minna JD, Scherle PA
    Cancer Cell, 2006;10(1):39-50.
    Species: Human
    Sample Types:
    Applications: Enzyme Assay
  14. Wnt5a signaling induces proliferation and survival of endothelial cells in vitro and expression of MMP-1 and Tie-2.
    Authors: Masckauchan TN, Agalliu D, Vorontchikhina M, Ahn A, Parmalee NL, Li CM, Khoo A, Tycko B, Brown AM, Kitajewski J
    Mol. Biol. Cell, 2006;17(12):5163-72.
    Species: Human
    Sample Types: Whole Cells
    Applications: Enzyme Assay
  15. Oxidized low-density lipoproteins stimulate extracellular matrix metalloproteinase Inducer (EMMPRIN) release by coronary smooth muscle cells.
    Authors: Haug C, Lenz C, Diaz F, Bachem MG
    Arterioscler. Thromb. Vasc. Biol., 2004;24(10):1823-9.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay

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Recombinant Human MMP-1 Protein, CF
By Anonymous on 11/09/2017
Application: Immunoassay Standard

The recombinant human MMP-1 was used to build an ELISA standard to measure MMP-1 in human serum and plasma samples.