MMP-9 Protein, Human (HEK293)

MMP-9 is an important member of the MMP protein family and regulates the extracellular matrix during physiological processes such as development and tissue remodeling. It involves arthritis and metastasis. MMP-9 Protein, Human (HEK293) is the recombinant human-derived MMP-9 protein, expressed by HEK293 , with tag free.

MMP-9, a member of the matrix metalloproteinase (MMP) family, plays a crucial role in modulating the extracellular matrix during various physiological processes, including embryonic development, reproduction, and tissue remodeling. These proteins are pivotal in both normal and pathological conditions, such as arthritis and metastasis. Typically secreted as inactive proproteins, MMPs are activated through cleavage by extracellular proteinases. The enzyme encoded by the MMP-9 gene specifically targets type IV and V collagens. Research in rhesus monkeys suggests its involvement in IL-8-induced mobilization of hematopoietic progenitor cells from the bone marrow, while murine studies propose a role in tumor-associated tissue remodeling. With biased expression observed in bone marrow (RPKM 144.9), lymph node (RPKM 49.8), and other tissues, MMP-9 emerges as a key player in the dynamic regulation of tissue architecture and cellular processes.

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 and the specific activity is >600 pmoles/min/μg. (Activation description: The proenzyme needs to be activated by APMA (HY-148905) for an activated form.)

• 
  Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2. The specific activity is 35188.587 pmol/min/µg, as measured under the described conditions.

Materials
Assay buffer：50 mM Tris,10 mM CaCl₂, 150 mM NaCl, 0.05% Brij-35(w/v), pH 7.5(TCNB)
MMP-9 Protein, Human (HEK293) (HY-P73300)
p-aminophenylmercuric acetate (APMA, HY-148905) , stored as a 100 mM solution diluted in DMSO.
Substrate：MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (HY-131498)
Standard: MCA-Pro-Leu-OH

Procedure
1. Standard Curve:Dilute standard in assay buffer to concentrations of 0, 1.5625, 3.125, 6.25, 12.5, 25, 50, and 100 μM. Add 100 μL of each dilution to a black microplate well. Measure fluorescence in kinetic mode for 5 minutes at excitation and emission wavelengths of 320 nm and 405 nm, respectively. Plot the measured RFU values on the y-axis and the standard concentrations on the x-axis to generate a standard curve and obtain the curve equation.
2. Dilute Human MMP-9 to 100 μg/mL in assay buffer.
3. Activate Human MMP-9 by adding APMA to a final concentration of 1 mM.
4. Incubate at 37°C for 24 hours.
5. Dilute the activated Human MMP-9 to 0.4 μg/mL in assay buffer.
6. Dilute the substrate to 20 μM in assay buffer.
7. Experimental group: Add 50 μL of 0.4 μg/mL Human MMP-9 to the plate and initiate the reaction by adding 50 μL of 20 μM substrate.
Control group: 50 μL of assay buffer and 50 μL of 20 μM substrate.
8. Read in kinetic mode for 5 minutes at excitation and emission wavelengths of 320 nm and 405 nm, respectively.
9. Calculate the specific activity.

Human

HEK293

Tag Free

NP_004985.2 (A20-D707)

4318  [NCBI]

Full Length of Mature Protein

APRQRQSTLVLFPGDLRTNLTDRQLAEEYLYRYGYTRVAEMRGESKSLGPALLLLQKQLSLPETGELDSATLKAMRTPRCGVPDLGRFQTFEGDLKWHHHNITYWIQNYSEDLPRAVIDDAFARAFALWSAVTPLTFTRVYSRDADIVIQFGVAEHGDGYPFDGKDGLLAHAFPPGPGIQGDAHFDDDELWSLGKGVVVPTRFGNADGAACHFPFIFEGRSYSACTTDGRSDGLPWCSTTANYDTDDRFGFCPSERLYTQDGNADGKPCQFPFIFQGQSYSACTTDGRSDGYRWCATTANYDRDKLFGFCPTRADSTVMGGNSAGELCVFPFTFLGKEYSTCTSEGRGDGRLWCATTSNFDSDKKWGFCPDQGYSLFLVAAHEFGHALGLDHSSVPEALMYPMYRFTEGPPLHKDDVNGIRHLYGPRPEPEPRPPTTTTPQPTAPPTVCPTGPPTVHPSERPTAGPTGPPSAGPTGPPTAGPSTATTVPLSPVDDACNVNIFDAIAEIGNQLYLFKDGKYWRFSEGRGSRPQGPFLIADKWPALPRKLDSVFEERLSKKLFFFSGRQVWVYTGASVLGPRRLDKLGLGADVAQVTGALRSGRGKMLLFSGRRLWRFDVKAQMVDPRSASEVDRMFPGVPLDTHDVFQYREKAYFCQDRFYWRVSSRSELNQVDQVGYVTYDILQCPED

76-95 kDa

• 
  Greater than 95% as determined by reducing SDS-PAGE.

Lyophilized powder

Lyophilized from a 0.22 μm filtered solution of 50 mM Tris, 0.15M NaCl, 0.01MCaCl₂, 0.05% Brij-35, pH 7.5 or 50 mM Tris, 150 mM NaCl, pH 7.5.
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.