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MMP-9 Protein, Human (HEK293)

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Based on 2 publication(s) in Google Scholar

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.

For research use only. We do not sell to patients.
  • Species: Human
  • Source: HEK293
  • Storage:
    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.
  • Experimental Validation Results for MMP-9 Protein, Human (HEK293)
  • Experimental Validation Results for MMP-9 Protein, Human (HEK293)
  • MMP-9 Protein, Human (HEK293) Product Image
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  • Biological Activity
  • Technical Parameters
  • Product Properties
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Biological Activity

Description

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.

Background

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.

Verified Bioactivity

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 and the specific activity is >1300 pmol/min/μg. (Activation description: The proenzyme needs to be activated by APMA (HY-148905) for an activated form.)

Assay Procedure

Materials
Assay buffer：50 mM Tris,10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35(w/v), pH 7.5(TCNB)
Test protein: MMP-9 Protein, Human (HEK293) (HY-P73300)
Activator: 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.

     Specific Activity (pmol/min/μg) =

Adjusted Vmax* (RFU/min) x Conversion Factor ** (pmol/RFU)
amount of enzyme (μg)

*Adjusted for Control
**Derived using calibration standard

Technical Parameters

  • Species Human
  • Source HEK293
  • Tag Tag Free
  • Accession

    NP_004985.2 (A20-D707)

  • Gene ID
    4318  [NCBI]
  • Molecular Construction
    • N-term
    • MMP-9 (A20-D707)
      Accession # NP_004985.2
    • C-term
  • Protein Length

    Full Length of Mature Protein (with Propeptide)

  • Synonyms

    MMP9; Matrix Metallopeptidase 9 (Gelatinase B, 92kDa Gelatinase, 92kDa Type IV Collagenase); Prev. CLG4B; Macrophage Gelatinase; Matrix Metalloproteinase 9 (Gelatinase B, 92kDa Gelatinase, 92kDa Type IV Collagenase); Type V Collagenase; Matrix Metalloproteinase-9; 92 KDa Gelatinase; MMP-9; Gelatinase B; GELB; MANDP2; Matrix Metallopeptidase 9; 92 KDa Type IV Collagenase

  • AA Sequence

    APRQRQSTLVLFPGDLRTNLTDRQLAEEYLYRYGYTRVAEMRGESKSLGPALLLLQKQLSLPETGELDSATLKAMRTPRCGVPDLGRFQTFEGDLKWHHHNITYWIQNYSEDLPRAVIDDAFARAFALWSAVTPLTFTRVYSRDADIVIQFGVAEHGDGYPFDGKDGLLAHAFPPGPGIQGDAHFDDDELWSLGKGVVVPTRFGNADGAACHFPFIFEGRSYSACTTDGRSDGLPWCSTTANYDTDDRFGFCPSERLYTQDGNADGKPCQFPFIFQGQSYSACTTDGRSDGYRWCATTANYDRDKLFGFCPTRADSTVMGGNSAGELCVFPFTFLGKEYSTCTSEGRGDGRLWCATTSNFDSDKKWGFCPDQGYSLFLVAAHEFGHALGLDHSSVPEALMYPMYRFTEGPPLHKDDVNGIRHLYGPRPEPEPRPPTTTTPQPTAPPTVCPTGPPTVHPSERPTAGPTGPPSAGPTGPPTAGPSTATTVPLSPVDDACNVNIFDAIAEIGNQLYLFKDGKYWRFSEGRGSRPQGPFLIADKWPALPRKLDSVFEERLSKKLFFFSGRQVWVYTGASVLGPRRLDKLGLGADVAQVTGALRSGRGKMLLFSGRRLWRFDVKAQMVDPRSASEVDRMFPGVPLDTHDVFQYREKAYFCQDRFYWRVSSRSELNQVDQVGYVTYDILQCPED

  • Molecular Weight

    Approximately 80-95 kDa, based on SDS-PAGE under reducing conditions, due to the glycosylation.

  • Glycosylation

    Yes

  • Purity

    ≥ 90%, as determined by reducing SDS-PAGE.

Product Properties

Appearance

Lyophilized powder

Formulation

1.Lyophilized from a 0.22 μm filtered solution of 50 mM Tris, 150 mM NaCl, 10 mM CaCl2, 0.05% Brij35, pH 7.5, 5% trehalose, 5% mannitol, 0.01% Tween 80.
2.Lyophilized from a 0.22 μm filtered solution of 50 mM Tris, 150 mM NaCl, pH 7.5.
3.Lyophilized from a 0.22 μm filtered solution of 50 mM Tris-HCl, 150 mM NaCl, pH 7.5.
4.Lyophilized from a 0.22 μm filtered solution of 50 mM Tris-HCl, 150 mM NaCl, pH 7.5, 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.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Reconstitution

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).

Storage & Stability

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.

Shipping

Room temperature in continental US; may vary elsewhere.

Calculators

Reconstitution Calculator

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration

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Dilution Calculator

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

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The Specific Activity Calculator Equation
  • Specific Activity (Unit/mg)
  • Biological Activity (ED50)

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)

Specific Activity (Unit/mg) Specific Activity (Unit/mg)
Unit/mg
= 106 ÷
Biological Activity (ED50) Biological Activity (ED50)
106 ÷
ng/mL
MOQ
Minimum order quantity
100 mg

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedChemExpress Validation
     ≥ 90%, as determined by reducing SDS-PAGE.

  • 1/16

    MedChemExpress Validation
     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.

  • MMP-9 Protein, Human (HEK293) Product Image
    1/16
  • Experimental Validation Results for MMP-9 Protein, Human (HEK293)
  • Experimental Validation Results for MMP-9 Protein, Human (HEK293)
  • MMP-9 Protein, Human (HEK293) Product Image