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.

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.

Recombinant Mouse MMP-9 Protein, CF

Name: Recombinant Mouse MMP-9 Protein, CF
Brand: R&D Systems
SKU: 909-MM
Rating: 5 (4 reviews)

Catalog #: 909-MM
19 Citations 4 Reviews Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
909-MM-010

Recombinant Mouse MMP-9 Protein Enzyme Activity

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All MMP-9 Proteins and Enzymes

Product Details

Citations (19)

FAQs

Supplemental Products

Reviews (4)

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

Recombinant Mouse MMP-9 Protein, CF Summary

Product Specifications

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

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-PLGL-Dpa-AR-NH₂ (Catalog # ES001). The specific activity is >1,500 pmol/min/µg, as measured under the described conditions.

Source

Mouse myeloma cell line, NS0-derived mouse MMP-9 protein
Ala20-Pro730

Accession #

P41245

N-terminal Sequence
Analysis

Ala20

Structure / Form

Pro form

Predicted Molecular Mass

78 kDa

SDS-PAGE

80-105 kDa, reducing conditions

Product Datasheets

909-MM

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.

909-MM

Formulation	Supplied as a 0.2 μm filtered solution in Tris, CaCl₂, NaCl 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% Brij-35 (w/v), pH 7.5 (TCNB)
Recombinant Mouse MMP-9 (rmMMP-9) (Catalog # 909-MM)
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

Dilute rmMMP-9 to 100 µg/mL in Assay Buffer.
Activate rmMMP-9 by adding APMA to a final concentration of 1 mM.
Incubate at 37 °C for 2 hours.
Dilute activated rmMMP-9 to 0.4 ng/µL in Assay Buffer.
Dilute Substrate to 20 µM in Assay Buffer.
Load into a black well plate 50 µL of the 0.4 ng/µL rmMMP-9 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 rmMMP-9.
Read at excitation 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:

rmMMP-9: 0.02 µg
Substrate: 10 µM

Scientific Data

Enzyme Activity

View Larger

Recombinant Mouse MMP-9 (Catalog # 909-MM) is measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 (Catalog # ES001).

Reconstitution Calculator

Background: MMP-9

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-9 (gelatinase B) can degrade a broad range of substrates including gelatin, collagen types IV and V, elastin and proteoglycan core protein. It is believed to act synergistically with interstitial collagenase (MMP-1) in the degradation of fibrillar collagens as it degrades their denatured gelatin forms. MMP-9 is produced by keratinocytes, monocytes, macrophages and PMN leukocytes. MMP-9 is present in most cases of inflammatory responses. Structurally, MMP-9 may be divided into five distinct domains: a pro-domain which is cleaved upon activation, a gelatin-binding domain consisting of three contiguous fibronectin type II units, a catalytic domain containing the zinc binding site, a proline-rich linker region, and a carboxyl terminal hemopexin-like domain. Compared to the Recombinant Human MMP‑9 (Catalog # 911-MP), the mouse enzyme contains extra sequences in the linker region and in the hemopexin-like domain, respectively.

Long Name

Matrix Metalloproteinase 9

Entrez Gene IDs

4318 (Human); 17395 (Mouse); 81687 (Rat); 102117693 (Cynomolgus Monkey)

Alternate Names

92 kDa gelatinase; 92 kDa type IV collagenase; CLG4B; EC 3.4.24; EC 3.4.24.35; Gelatinase B; GELB; macrophage gelatinase; MANDP2; matrix metallopeptidase 9; matrix metalloproteinase 9; matrix metalloproteinase-9; MMP9; MMP-9; type V collagenase

Related Research Areas

Citations for Recombinant Mouse MMP-9 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.

19 Citations: Showing 1 - 10
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Secretomics reveals gelatinase substrates at the blood-brain barrier that are implicated in astroglial barrier function
Authors: Burmeister, M;Fraunenstein, A;Kahms, M;Arends, L;Gerwien, H;Deshpande, T;Kuhlmann, T;Gross, CC;Naik, VN;Wiendl, H;Klingauf, J;Meissner, F;Sorokin, L;
Science advances
Species: Mouse
Sample Types: Recombinant Protein
Applications: Bioassay
Carnosine Protects against Cerebral Ischemic Injury by Inhibiting Matrix-Metalloproteinases
Authors: EH Kim, ES Kim, D Shin, D Kim, S Choi, YJ Shin, KA Kim, D Noh, AB Caglayan, GK Rajanikant, A Majid, ON Bae
International Journal of Molecular Sciences, 2021-07-13;22(14):.
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay
Loss of IL-33 enhances elastase-induced and cigarette smoke extract-induced emphysema in mice
Authors: D Morichika, A Taniguchi, N Oda, U Fujii, S Senoo, J Itano, A Kanehiro, Y Kitaguchi, M Yasuo, M Hanaoka, T Satoh, S Akira, K Kiura, Y Maeda, N Miyahara
Respiratory Research, 2021-05-15;22(1):150.
Species: Mouse
Sample Types: BALF
Applications: Reference Standard
Blood substitution therapy rescues the brain of mice from ischemic damage
Authors: X Ren, H Hu, I Farooqi, JW Simpkins
Nat Commun, 2020-08-25;11(1):4078.
Species: Mouse
Sample Types: In Vivo
Applications: In Vivo
Matrix metalloproteinase 9 facilitates Zika virus invasion of the testis by modulating the integrity of the blood-testis barrier.
Authors: L Hui, Y Nie, S Li, M Guo, W Yang, R Huang, J Chen, Y Liu, X Lu, Z Chen, Q Yang, Y Wu
PLoS Pathog, 2020-04-17;16(4):e1008509.
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay
Tissue Inhibitor of Metalloproteinase-1 Promotes Polymorphonuclear Neutrophil (PMN) Pericellular Proteolysis by Anchoring Matrix Metalloproteinase-8 and -9 to PMN Surfaces
Authors: X Wang, J Rojas-Quin, J Wilder, Y Tesfaigzi, D Zhang, CA Owen
J. Immunol., 2019-04-24;0(0):.
Species: Transgenic Mouse
Sample Types: Whole Cells
Applications: Bioassay
Dentin Sialoprotein is a Novel Substrate of Matrix Metalloproteinase 9 in vitro and in vivo
Authors: G Yuan, L Chen, J Feng, G Yang, Q Ni, X Xu, C Wan, M Lindsey, KJ Donly, M MacDougall, Z Chen, S Chen
Sci Rep, 2017-02-14;7(0):42449.
Species: Mouse
Sample Types: Recombinant Protein
Applications: Zymography
Accelerated biodegradation of silk sutures through matrix metalloproteinase activation by incorporating 4-hexylresorcinol
Authors: YY Jo, H Kweon, DW Kim, MK Kim, SG Kim, JY Kim, WS Chae, SP Hong, YH Park, SY Lee, JY Choi
Sci Rep, 2017-02-13;7(0):42441.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay
Reversal of TREM-1 ectodomain shedding and improved bacterial clearance by intranasal metalloproteinase inhibitors
Mucosal Immunol, 2016-12-14;0(0):.
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay
Neuropeptide Y induces hematopoietic stem/progenitor cell mobilization by regulating matrix metalloproteinase-9 activity through Y1 receptor in osteoblasts
Authors: MH Park, JK Lee, N Kim, WK Min, JE Lee, KT Kim, H Akiyama, H Herzog, EH Schuchman, HK Jin, JS Bae
Stem Cells, 2016-05-18;0(0):.
Species: Mouse
Sample Types: In Vivo
Applications: In Vivo

G protein-coupled receptor kinase 6 deficiency promotes angiogenesis, tumor progression, and metastasis.
Authors: Raghuwanshi S, Smith N, Rivers E, Thomas A, Sutton N, Hu Y, Mukhopadhyay S, Chen X, Leung T, Richardson R
J Immunol, 2013-04-15;190(10):5329-36.
Species: Mouse
Sample Types: Recombinant Protein, Tissue Homogenates
Applications: ELISA (Standard), Western Blot

Silibinin inhibits TPA-induced cell migration and MMP-9 expression in thyroid and breast cancer cells.
Authors: Oh S, Jung S, Han J, Kim S, Kim J, Nam S, Lee J, Kim J
Oncol Rep, 2013-01-24;29(4):1343-8.

In vivo processing of CXCL5 (LIX) by matrix metalloproteinase (MMP)-2 and MMP-9 promotes early neutrophil recruitment in IL-1beta-induced peritonitis.
Authors: Song J, Wu C, Zhang X, Sorokin L
J Immunol, 2012-12-07;190(1):401-10.
Species: Mouse
Sample Types: Whole Cells
Applications: Cell Culture

Ocular neovascularization caused by herpes simplex virus type 1 infection results from breakdown of binding between vascular endothelial growth factor A and its soluble receptor.
Authors: Suryawanshi A, Mulik S, Sharma S
J. Immunol., 2011-02-16;186(6):3653-65.
Species: Mouse
Sample Types: Cell Lysates
Applications: Enzyme Assay

The selective MMP-12 inhibitor, AS111793 reduces airway inflammation in mice exposed to cigarette smoke.
Authors: Le Quement C, Guenon I, Gillon JY, Valenca S, Cayron-Elizondo V, Lagente V, Boichot E
Br. J. Pharmacol., 2008-05-19;154(6):1206-15.
Applications: Zymography

A novel proteolytic cascade generates an extracellular matrix-derived chemoattractant in chronic neutrophilic inflammation.
Authors: Gaggar A, Jackson PL, Noerager BD, O'Reilly PJ, McQuaid DB, Rowe SM, Clancy JP, Blalock JE
J. Immunol., 2008-04-15;180(8):5662-9.
Species: Mouse
Sample Types: In Vivo
Applications: In Vivo

A GMCSF and IL-15 fusokine leads to paradoxical immunosuppression in vivo via asymmetrical JAK/STAT signaling through the IL-15 receptor complex.
Authors: Rafei M, Wu JH, Annabi B, Lejeune L, Francois M, Galipeau J
Blood, 2006-11-02;109(5):2234-42.
Applications: Western Blot

Activity-dependent release of precursor nerve growth factor, conversion to mature nerve growth factor, and its degradation by a protease cascade.
Authors: Bruno MA, Cuello AC
Proc. Natl. Acad. Sci. U.S.A., 2006-04-17;103(17):6735-40.
Species: Rat
Sample Types: Tissue Homogenates
Applications: Bioassay

Angiotensin II-accelerated atherosclerosis and aneurysm formation is attenuated in osteopontin-deficient mice.
Authors: Bruemmer D, Collins AR, Noh G, Wang W, Territo M, Arias-Magallona S, Fishbein MC, Blaschke F, Kintscher U, Graf K, Law RE, Hsueh WA
J. Clin. Invest., 2003-11-01;112(9):1318-31.
Applications: Zymography

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.
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.