Recombinant Human MMP-9 (NS0-expressed) Protein, CF

Name: Recombinant Human MMP-9 (NS0-expressed) Protein, CF
Brand: R&D Systems
SKU: 911-MPN

Catalog #: 911-MPN
2 Citations Datasheet / COA / SDS

Newer Version Available: 911-MP

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Discontinued Product

911-MPN has been discontinued and is replaced by 911-MP.

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Citations (2)

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

Recombinant Human MMP-9 (NS0-expressed) 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 the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH₂ (Catalog # ES001). The specific activity is >1,300 pmol/min/µg measured under the described conditions.

Source

Mouse myeloma cell line, NS0-derived human MMP-9 protein
Ala20-Asp707(Gln279Arg)

Accession #

P14780

N-terminal Sequence
Analysis

Ala20

Structure / Form

Pro form

Predicted Molecular Mass

76 kDa

SDS-PAGE

97 kDa, reducing conditions

Product Datasheets

911-MPN

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.

911-MPN

Formulation	Supplied as a 0.2 μm filtered solution in Tris-HCl, 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% Brij-35 (w/v), pH 7.5 (TCNB)
Recombinant Human MMP-9 (rhMMP-9) (Catalog # 911-MPN)
p-aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563), prepare a 100 mM stock in DMSO
Substrate: MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH₂ (Catalog # ES001)
F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent

Dilute rhMMP-9 to 100 µg/mL in Assay Buffer.
Activate rhMMP-9 by adding APMA to a final concentration of 1 mM.
Incubate at 37 °C for 24 hours.
Dilute activated rhMMP-9 to 0.2 ng/µL in Assay Buffer.
Dilute Substrate to 20 µM in Assay Buffer.
Load 50 µL of the 0.2 ng/µL rhMMP-9 into a plate and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 20 µM Substrate.
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:

rhMMP-9: 0.010 µg
Substrate: 10 µM

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 maybe 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. In addition to the human enzyme, the recombinant mouse MMP-9 is also available (Catalog # 909-MM).

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 Human MMP-9 (NS0-expressed) 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.

2 Citations: Showing 1 - 2
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Matrix metalloproteinase (MMP)-2, MMP-9, semen quality and sperm longevity in fractionated stallion semen
Authors: M Kareskoski, J Vakkamäki, K Laukkanen, M Palviainen, A Johannisso, T Katila
Theriogenology, 2021-02-02;164(0):93-99.
Species: Equine
Sample Types: Semen
Applications: Gelatin Zymography Control
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