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

Name: Recombinant Human MMP-1 Protein, CF
Brand: R&D Systems
SKU: 901-MP
Rating: 4.5 (2 reviews)

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.

Recombinant Human MMP-1 Protein, CF

Catalog #: 901-MP
16 Citations 2 Reviews Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
901-MP-010

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Citations (16)

FAQs

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Reviews (2)

Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human MMP-1 Protein, CF Summary

Product Specifications

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 #

P03956

N-terminal Sequence
Analysis

Phe20

Structure / Form

Pro form

Predicted Molecular Mass

52 kDa

SDS-PAGE

52-55 kDa, reducing conditions

Product Datasheets

901-MP

Product Datasheet

COA

SDS

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

Dilute rhMMP-1 to 50 µg/mL in Assay Buffer.
Activate 50 µg/mL rhMMP-1 by adding APMA to a final concentration of 1 mM.
Incubate at 37 °C for 2 hours.
Dilute activated rhMMP-1 to 1 ng/µL in Assay Buffer.
Dilute Substrate to 20 µM in Assay Buffer.
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.
Read at excitiation and emission wavelengths of 320 nm and 405 nm, respectively, in kinetic mode for 5 minutes.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (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

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

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

Related Research Areas

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.

16 Citations: Showing 1 - 10
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MMP-1 promotes osteogenic differentiation of human bone marrow mesenchymal stem cells via the JNK and ERK pathway
Authors: Y Wu, Y Tang, X Zhang, Z Chu, Y Liu, C Tang
Int J Biochem Cell Biol, 2020-11-04;129(0):105880.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay
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., 2018-04-04;0(0):.
Applications: Enzyme Assay
MMP-1 Activation Contributes to Airway Smooth Muscle Growth and Asthma Severity
Authors: Shams-Un-Nisa Naveed
Am. J. Respir. Crit. Care Med, 2017-04-15;0(0):.
Species: Human
Sample Types: Protein
Applications: Bioassay
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, 2017-03-07;19(4):271-278.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay
Transformed MDCK cells secrete elevated MMP1 that generates LAMA5 fragments promoting endothelial cell angiogenesis
Authors: Shashi K Gopal
Sci Rep, 2016-06-21;6(0):28321.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay
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, 2014-04-23;192(11):4977-88.
Species: Human
Sample Types: Cell Culture Supernates
Applications: Bioassay
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, 2013-06-19;288(31):22821-35.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay
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, 2013-02-05;54(2):1014-25.
Species: Bovine
Sample Types: Protein
Applications: Enzyme Assay
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
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, 2012-02-17;378(1):72-80.
Applications: ELISA (Standard)

Development and validation of sandwich ELISA microarrays with minimal assay interference.
Authors: Gonzalez RM, Seurynck-Servoss SL, Crowley SA
J. Proteome Res., 2008-04-19;7(6):2406-14.
Applications: ELISA (Standard)

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., 2007-12-11;193(1):140-8.
Species: Rat
Sample Types: Recombinant Protein
Applications: Enzyme Assay

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), 2007-01-11;46(5):752-8.
Applications: ELISA (Standard)

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, 2006-10-11;17(12):5163-72.
Species: Human
Sample Types: Whole Cells
Applications: Enzyme Assay

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, 2006-07-01;10(1):39-50.
Species: Human
Sample Types:
Applications: Enzyme Assay

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., 2004-08-19;24(10):1823-9.
Species: Human
Sample Types: Protein
Applications: Bioassay

FAQs

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.
Do I need to add zinc to this assay buffer?
- In our experience, assaying the activity of this MMP with a fluorogenic substrate does not require the addition of zinc to the assay buffer.
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.
Does this MMP enzyme need to be activated to work?
- Yes, this enzyme requires activation prior to use.
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.