Recombinant Human MMP-9 Activated Protein, CF New

Catalog #: 11602-MP Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
11602-MP-020

Recombinant Human MMP‑9 Activated Protein Enzyme Activity.

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

Recombinant Human MMP-9 Activated Protein, CF Summary

Product Specifications

Endotoxin Level

<0.10 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 >1000 pmol/min/μg, as measured under the described conditions.

Source

Chinese Hamster Ovary cell line, CHO-derived human MMP-9 protein
Ala20-Asp707 (Gln279Arg)
The proform was activated.

Accession #

P14780.2

N-terminal Sequence
Analysis

Phe107 & Gly547

Structure / Form

Activated

SDS-PAGE

60-85, 38-42, 16-21 kDa, under reducing conditions

Product Datasheets

11602-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.

11602-MP

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 Human MMP-9 Activated (rhMMP-9) (Catalog # 11602-MP)
Substrate: MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES001)
Black 96-well plate
Plate Reader with Fluorescence Read Capability

Dilute rhMMP-9 to 0.4 µg/mL in Assay Buffer.
Dilute Substrate to 20 µM in Assay Buffer.
Load 50 µL of 0.4 µg/mL 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.

Per Well:

rhMMP-9: 0.020 μg
Substrate: 10 µM

Scientific Data

Enzyme Activity

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Recombinant Human MMP-9 Activated (Catalog # 11602-MP) is measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 (ES001).

Reconstitution Calculator

Background: MMP-9

Recombinant human Active Matrix Metalloproteinase 9 (MMP-9), also known as gelatinase B, is a member of the MMP family of zinc and calcium-dependent endopeptidases. MMP-9 protein is synthesized as a pre-proenzyme in specific cells such as neutrophils, macrophages, fibroblasts and endothelial cells (1). Expressed MMP-9 contains a signal peptide to transport it to the extracellular matrix (ECM), a hinge region, a propeptide region, a catalytic domain, and a hemopexin-like domain that is important for substrate recognition (2, 3). In addition to having three fibronectin type II domains that contribute to substrate binding and an active site, the catalytic domain of MMP-9 also contains a zinc-binding region that interacts with a cysteine in the propeptide to maintain latency. Consequently, removal of the propeptide through cleavage by proteases in the ECM, such as MMP-3, activates the protein (2, 3). MMP-9 has specificity for targets containing an established preferred consensus sequence (3-5) and has a broad range of substrates within the ECM including gelatin, collagen, elastin that contributes to its role in ECM remodeling and extracellular domain cell surface protein release from the plasma membrane (3). Due to its activity in the ECM, MMP-9 plays a role in many biological processes and can serve as a biomarker in tumor invasion and mestastasis (3, 6) of many cancers including colon, ovarian, breast, osteosarcoma, and lung cancers (7-11) making it a therapeutic target (6, 8, 12). MMP-9 modeling of the ECM also leads to a pivotal role in other inflammation- and autoimmune-related diseases (3, 13, 14).

References

Vandooren, J. et al. (2013) Crit. Rev. Biochem. Mol. Biol. 48: 222.
Roeb, E. et al. (2002) J. Biol. Chem. 277: 50326.
Huang, H. (2018) Sensors. 18:3249.
Kridel, S.J. et al. (2001) J. Biol. Chem. 276: 20572.
Prudova, A. et al. (2010) Mol. Cell Proteom. 9:894.
Kalali, D. (2023) Glob. Med. Genet. 10:48.
Hu, X. et al. (2012) Arch. Glynecol. Obstet. 286:1537.
Wang, J. et al. (2014) Clin. Chim. Acta. 433:225.
Blanco-Prieto, S. et al. (2017) BMC Cancer 17:823.
Liang, S. and L. Chang (2018) Biomark. Med. 12:393.
Malik, S. et al. (2024) Sci. Rep. 14:15117.
Roy, R. et al. (2009) J. Clin. Oncol. 27:5287.
Ram, M. et al. (2006) J. Clin. Immunol. 26:299.
Kim, I.S. et al. (2023) Curr. Med. Chem. 30:2075.

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