MMP-1 Protein, Human (HEK293, C-His)
Based on 1 Customer Validation
The MMP-1 protein acts as an enzyme capable of cleaving types I, II, and III collagen at specific sites within the helical domain. In addition, it exhibits lytic activity against type VII and type X collagen. MMP-1 Protein, Human (HEK293, C-His) is the recombinant human-derived MMP-1, expressed by HEK293, with C-6*His labeled tag.
- Species: Human
- Source: HEK293
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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.
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Biological Activity
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Technical Parameters
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Product Properties
- Documentation
- Help & FAQs
Biological Activity
Description
The MMP-1 protein acts as an enzyme capable of cleaving types I, II, and III collagen at specific sites within the helical domain. In addition, it exhibits lytic activity against type VII and type X collagen. MMP-1 Protein, Human (HEK293, C-His) is the recombinant human-derived MMP-1, expressed by HEK293, with C-6*His labeled tag.
Background
Matrix metalloproteinase-1 (MMP-1) is a proteolytic enzyme known for its role in extracellular matrix remodeling. It exhibits a broad substrate specificity, cleaving collagens of types I, II, and III at a specific site within the helical domain. Additionally, MMP-1 demonstrates activity against collagens of types VII and X. Notably, in the context of HIV infection, MMP-1 interacts with and cleaves the secreted viral Tat protein, resulting in a decrease in neuronal Tat-mediated neurotoxicity. This interaction underscores the diverse functions of MMP-1 beyond its canonical role in extracellular matrix degradation, implicating it in processes associated with viral infection and neuroprotection (
Verified Bioactivity
Measured by its ability to cleave 20 µM fluorogenic peptide substrate, Mca-KPLGL-Dpa-AR-NH2 that incubate at room temperature in kinetic mode for 5 minutes. The specific activity is >4000 pmol/min/µg, as measured under the described conditions. (Activation description: The proenzyme needs to be activated by APMA (HY-148905) for an activated form.)
MCE Validation Data
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Purity - SDS-PAGE
Purity - SDS-PAGE
Assay Procedure
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-1 Protein, Human (HEK293, C-His) (HY-P70250A)
Activator: 4-Aminophenylmercuriacetic acid (APMA, HY-148905), dissolved in DMSO to 100 mM
Substrate: MCA-Lys-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2< (HY-131498)
Standard: MCA-Pro-Leu-OH
Procedure
1. Standard curve: Dilute MCA-Pro-Leu-OH to concentrations of 0, 1.5625, 3.125, 6.25, 12.5, 25, 50, and 100 μmol/L using assay buffer. Add 100 μL of each dilution to black microplate wells. Read at 320 nm excitation and 405 nm emission (top read) in kinetic mode for 5 minutes. Plot the standard curve with measured RFU on the y-axis and standard substance amount on the x-axis to derive the standard curve equation.
2. Resuspend proteins in water to 200 μg/mL. Dilute Human MMP-1 to 50 μg/mL using assay buffer containing 1 mM APMA.
3. Activation: Incubate at 37°C for 2 hours.
4. Dilute the activated Human MMP-1 protein to 1 ng/μL using buffer.
5. Dilute the substrate to 20 μM using assay buffer.
6. Experimental group: 50 μL, 1 ng/μL Human MMP-1 protein + 50 μL, 20 μM substrate.
Control group: 50 μL, 20 μM substrate + 50 μL buffer.
7. Read for 5 minutes in kinetic mode at excitation and emission wavelengths of 320 nm and 405 nm respectively.
8. Calculate specific activity:
Specific Activity (pmol/min/μg) = | Adjusted Vmax* (RFU/min) x Conversion Factor ** (pmol/RFU) |
| amount of enzyme (μg) |
*Adjusted for Substrate Blank
**Derived using calibration standard
Technical Parameters
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Species Human
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Source HEK293
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Tag C-6*His
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Accession
P03956 (F20-N469)
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Gene ID4312
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Protein Length
Full Length of Mature Protein (with propeptide)
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Synonyms
MMP1; Matrix Metallopeptidase 1 (Interstitial Collagenase); Prev. CLG; Matrix Metalloproteinase 1; Interstitial Collagenase; Matrix Metalloprotease 1; Fibroblast Collagenase; Alternative Protein MMP1; Matrix Metalloproteinase 1 (Interstitial Collagenase);
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AA Sequence
FPATLETQEQDVDLVQKYLEKYYNLKNDGRQVEKRRNSGPVVEKLKQMQEFFGLKVTGKPDAETLKVMKQPRCGVPDVAQFVLTEGNPRWEQTHLTYRIENYTPDLPRADVDHAIEKAFQLWSNVTPLTFTKVSEGQADIMISFVRGDHRDNSPFDGPGGNLAHAFQPGPGIGGDAHFDEDERWTNNFREYNLHRVAAHELGHSLGLSHSTDIGALMYPSYTFSGDVQLAQDDIDGIQAIYGRSQNPVQPIGPQTPKACDSKLTFDAITTIRGEVMFFKDRFYMRTNPFYPEVELNFISVFWPQLPNGLEAAYEFADRDEVRFFKGNKYWAVQGQNVLHGYPKDIYSSFGFPRTVKHIDAALSEENTGKTYFFVANKYWRYDEYKRSMDPGYPKMIAHDFPGIGHKVDAVFMKDGFFYFFHGTRQYKFDPKTKRILTLQKANSWFNCRKN
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Molecular Weight
Approximately 51-61 kDa, based on SDS-PAGE under reducing conditions, due to the glycosylation.
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Purity
≥ 95%, as determined by reducing SDS-PAGE.
Product Properties
Lyophilized powder
1.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4.
2.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 8% trehalose.
<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.
Documentation
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Data Sheet (238 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Handling Instructions (2659 KB)
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
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