MMP-2 Protein, Human (HEK293)

MMP-2 protein is a multifunctional metalloproteinase that actively participates in physiological processes such as vascular remodeling, angiogenesis, tissue repair, tumor invasion, inflammation, and atherosclerotic plaque rupture. In addition to degrading extracellular matrix proteins, it also acts on non-matrix proteins to promote vasoconstriction. MMP-2 Protein, Human (HEK293) is the recombinant human-derived MMP-2 protein, expressed by HEK293 , with tag free.

The MMP-2 protein, a ubiquitinous metalloproteinase, actively participates in a spectrum of physiological processes, including vasculature remodeling, angiogenesis, tissue repair, tumor invasion, inflammation, and atherosclerotic plaque rupture. Beyond its role in degrading extracellular matrix proteins, this protein demonstrates versatility by acting on non-matrix proteins, such as big endothelial 1 and beta-type CGRP, thereby promoting vasoconstriction. Additionally, it cleaves KISS at a Gly-|-Leu bond and appears to play a role in myocardial cell death pathways. By regulating the activity of GSK3beta and cleaving GSK3beta in vitro, it contributes to myocardial oxidative stress. In association with MMP14, MMP-2 is involved in the formation of fibrovascular tissues. Notably, the C-terminal non-catalytic fragment of MMP-2, known as PEX, possesses anti-angiogenic and anti-tumor properties, inhibiting cell migration and adhesion to FGF2 and vitronectin. Furthermore, it serves as a ligand for integrin alpha-v/beta3 on the surface of blood vessels.

1. Measured by its ability to cleave the fluorogenic peptide substrate Mca-PLGL-Dpa-AR-NH2 ((HY-131498) and the specific activity is > 1,000 pmoles/min/µg. (Activation description: The proenzyme needs to be activated by APMA (HY-148905) for an activated form.)
2. Measured by its binding ability in a functional ELISA. Immobilized Human MMP-2 at 2 μg/mL (100 μl/well) can bind Human TIMP2 hFc and the EC₅₀ is 6.0-30.0 ng/mL.

• 
  Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2. The specific activity is 11011 pmol/min/µg, as measured under the described conditions.

Materials
Assay buffer: 50 mM Tris, 10 mM CaCl₂, 150 mM NaCl, 0.05% (w/v) Brij 35, pH 7.5 (TCNB)
Test protein: MMP-2 Protein, Human (HEK293) (HY-P73296)
Substrate: MOCAc-PLGL(Dpa) AR (HY-131498)
Standard：MCA-Pro-Leu-OH
Activator: p-aminophenylmercuric acetate (APMA), dissolved in DMSO to 100 mM
F16 Black Maxisorp Plate

Procedure
1. Dilute Human MMP-2 to 100 μg/mL in assay buffer.
2. Activate Human MMP-2 by adding APMA to a final concentration of 1 mM in assay buffer.
3. Incubate at 37°C for 1 hour.
4. Dilute the activated Human MMP-2 to 0.2 μg/mL in assay buffer.
5. Dilute the substrate to 20 μM in assay buffer.
6. Experimental group: 50 μL of different concentrations of Human MMP-2 protein + 50 μL of 20 μM substrate.
7. Control group: 50 μL of 20 μM substrate + 50 μL of buffer.
8. Read in kinetic mode for 5 minutes at excitation and emission wavelengths of 320 nm and 405 nm, respectively.
9. Calculate Specific Activity:

Human

HEK293

Tag Free

P08253-1/NP_004521.1 (A30-C660)

4313  [NCBI]

Full Length of Isoform-1 Mature Protein

APSPIIKFPGDVAPKTDKELAVQYLNTFYGCPKESCNLFVLKDTLKKMQKFFGLPQTGDLDQNTIETMRKPRCGNPDVANYNFFPRKPKWDKNQITYRIIGYTPDLDPETVDDAFARAFQVWSDVTPLRFSRIHDGEADIMINFGRWEHGDGYPFDGKDGLLAHAFAPGTGVGGDSHFDDDELWTLGEGQVVRVKYGNADGEYCKFPFLFNGKEYNSCTDTGRSDGFLWCSTTYNFEKDGKYGFCPHEALFTMGGNAEGQPCKFPFRFQGTSYDSCTTEGRTDGYRWCGTTEDYDRDKKYGFCPETAMSTVGGNSEGAPCVFPFTFLGNKYESCTSAGRSDGKMWCATTANYDDDRKWGFCPDQGYSLFLVAAHEFGHAMGLEHSQDPGALMAPIYTYTKNFRLSQDDIKGIQELYGASPDIDLGTGPTPTLGPVTPEICKQDIVFDGIAQIRGEIFFFKDRFIWRTVTPRDKPMGPLLVATFWPELPEKIDAVYEAPQEEKAVFFAGNEYWIYSASTLERGYPKPLTSLGLPPDVQRVDAAFNWSKNKKTYIFAGDKFWRYNEVKKKMDPGFPKLIADAWNAIPDNLDAVVDLQGGGHSYFFKGAYYLKLENQSLKSVKFGSIKSDWLGC

Approximately 72-74 kDa, based on SDS-PAGE under reducing conditions, due to the glycosylation.

• 
  ≥ 95%, as determined by reducing SDS-PAGE.

Lyophilized powder

1.Lyophilized from a 0.22 μm filtered solution of 0.05 % Brij-35, 150 mM NaCl, 5 mM CaCl₂, 50 mM Tris, pH 7.5.
2.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 8% trehalose.
Please refer to the lot-specific COA for specific buffer information.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.