Recombinant Human MMP-1

Recombinant Human MMP-1 (Matrix Metalloproteinase-1) is a valuable tool for research applications due to its well-characterized enzymatic activity and diverse biological roles. Here are several reasons why you should consider using Recombinant Human MMP-1 in your research:

1. Defined and Consistent Activity

Recombinant Human MMP-1 is produced under controlled conditions, ensuring high purity (>90–95%) and batch-to-batch consistency. This allows for reproducible experimental results, which is critical for enzyme assays, biochemical studies, and functional investigations.

2. Versatile Applications

Recombinant MMP-1 is suitable for a wide range of research applications, including:

• Enzyme activity assays to study collagen degradation and substrate specificity.
• Cell culture studies to investigate the effects of MMP-1 on cell signaling, migration, and morphology.
• ELISA standard for quantifying MMP-1 levels in biological samples.
• Structural studies (e.g., crystallography) to understand the active conformation of MMP-1.

3. Role in Extracellular Matrix Remodeling

MMP-1 is a key enzyme in the degradation of fibrillar collagens (types I, II, and III), which are major components of the extracellular matrix. This makes it essential for studies on tissue remodeling, wound healing, fibrosis, and cancer metastasis.

4. Involvement in Cell Signaling and Plasticity

Recent research shows that MMP-1 can activate cell surface receptors such as Protease-Activated Receptor 1 (PAR1), influencing intracellular signaling pathways, neuronal plasticity, dendritic complexity, and even behavioral phenotypes. This expands its utility beyond matrix degradation to studies on cell signaling and neurobiology.

5. Relevance to Disease Mechanisms

MMP-1 is implicated in various pathological conditions, including:

• Cancer progression and metastasis (due to its ability to degrade ECM and facilitate tumor cell invasion).
• Inflammatory diseases (such as arthritis and asthma).
• Neurological disorders (where MMP-1 may affect synaptic plasticity and glial activation).

6. Availability in Multiple Formats

Recombinant Human MMP-1 is available in various forms (e.g., carrier-free, His-tagged, active full-length protein) and expression systems (E. coli, HEK 293, mouse myeloma), allowing you to select the most appropriate product for your experimental needs.

7. Facilitates Mechanistic Studies

Using recombinant MMP-1 allows you to directly test the effects of the enzyme on specific substrates, cells, or signaling pathways, helping to elucidate molecular mechanisms in both physiological and pathological contexts.

In summary, Recombinant Human MMP-1 is a reliable, versatile, and biologically relevant reagent that can advance your research in areas such as extracellular matrix biology, cell signaling, disease mechanisms, and therapeutic development.

Yes, recombinant human MMP-1 can be used as a standard for quantification and calibration in ELISA assays, though there are important considerations regarding its specific form and intended application.

Suitability as ELISA Standards

Recombinant human MMP-1 proteins are specifically designed and validated for use as ELISA standards. Many commercial ELISA kits employ NS0-expressed recombinant human MMP-1 as their calibration standards, and natural human MMP-1 shows dose-response curves that are parallel to standard curves obtained using recombinant standards, indicating reliable quantification.

Critical Distinction: Pro-MMP-1 vs. Active MMP-1

An essential consideration is the activation state of the recombinant protein. The antibodies in many ELISA kits specifically recognize pro-MMP-1 (the inactive zymogen form) and will not cross-react with active MMP-1. If your assay is designed to measure pro-MMP-1, you must use recombinant pro-MMP-1 as your standard. Conversely, if measuring total or active MMP-1, you need the appropriate form of recombinant protein.

Formulation Considerations

When selecting recombinant MMP-1 for use as an ELISA standard, consider the carrier protein formulation:

• With BSA (bovine serum albumin): Recommended for use in cell or tissue culture applications and as ELISA standards
• Carrier-free formulations: Better suited for specific applications where carrier proteins may interfere

Note that ELISA standard recombinant proteins are not typically tested for bioassay applications, so if you require the protein for functional enzyme assays, verify that your recombinant preparation has been validated for that purpose.

Validation Requirements

Before using recombinant MMP-1 as your standard, ensure that:

• The recombinant protein matches the target form (pro-MMP-1, active MMP-1, or total MMP-1) that your assay is designed to measure
• You prepare diluted standards within 60 minutes of preparation for optimal stability
• You account for potential variations in diluent, pipetting technique, incubation conditions, and kit age, as these can affect binding and standard curve generation

Recombinant Human MMP-1 has been validated for several key applications in published research:

Primary Research Applications

Enzymatic Activity Assays

Recombinant MMP-1 is extensively used to characterize enzymatic activity in vitro. The protein demonstrates robust proteolytic activity, cleaving specific peptide substrates such as Mca-KPLGL-Dpa-AR-NH2 with activity levels exceeding 200 pmol/min/µg. This application is fundamental for studying the enzyme's kinetic properties and substrate specificity.

Bioactivity and Bioassay Studies

The protein has been validated for bioassay applications to assess its functional capacity in biological systems. These studies examine how recombinant MMP-1 performs in cell and tissue culture contexts, providing insights into its physiological roles and interactions with cellular components.

Neutralization Studies

Recombinant MMP-1 serves as a tool for neutralization assays, where researchers evaluate the effectiveness of inhibitors, antibodies, or other blocking agents against the enzyme's activity. This application is critical for drug development and therapeutic target validation.

Immunoassay Applications

The protein functions as a standard in ELISA and other immunoassay formats, enabling quantitative measurement of endogenous MMP-1 levels in biological samples and facilitating standardization across different experimental protocols.

Research Context

These validated applications support investigations into MMP-1's diverse biological roles, including extracellular matrix remodeling through collagen degradation, enzyme cascade regulation, cytokine modulation, and cell surface molecule interactions. The protein has been employed in studies examining MMP-1's involvement in pathological processes such as arthritis, cancer metastasis, atherosclerosis, and periodontal disease.

To reconstitute and prepare Recombinant Human MMP-1 protein for cell culture experiments, dissolve the lyophilized protein in sterile buffer (commonly PBS, pH 7.4, or sterile distilled water) to a final concentration of 0.1–1.0 mg/mL. Avoid vortexing or vigorous pipetting to prevent protein denaturation.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all protein is at the bottom.
• Add sterile buffer (e.g., 10 mM PBS, pH 7.4, or sterile distilled water) to achieve the desired concentration (typically 0.1–1.0 mg/mL for stock solutions).
• Gently mix by slow pipetting or gentle inversion. Do not vortex or shake vigorously.
• Let the solution sit at room temperature for 15–30 minutes to fully dissolve the protein.
• Aliquot the reconstituted protein to minimize freeze-thaw cycles, which can degrade activity.
• For long-term storage, add a carrier protein or stabilizer (e.g., 0.1% BSA, 5% HSA, 10% FBS, or 5% trehalose) if compatible with your downstream application.
• Store aliquots at –20°C or –80°C. Avoid repeated freeze/thaw cycles.

Preparation for cell culture experiments:

• Dilute the stock solution to the working concentration required for your experiment using cell culture medium or assay buffer. Typical working concentrations range from nanograms to micrograms per milliliter, depending on the assay design and cell type.
• If activation of MMP-1 is required (e.g., for enzymatic assays), incubate with 1 mM APMA (amino-phenyl mercuric acetate) at 37°C for 1–2 hours. For cell culture, activation may not be necessary unless specifically required by your experimental design.
• Filter-sterilize the final working solution if sterility is critical for cell culture.

Additional notes:

• Always consult the specific product datasheet for recommended reconstitution buffer and concentration, as formulations may vary.
• Avoid foaming and excessive agitation, which can denature the protein.
• For cell-based assays, confirm that any additives (e.g., carrier proteins, APMA) are compatible with your cells and experimental goals.

Summary of key points:

• Use sterile PBS (pH 7.4) or distilled water for reconstitution.
• Final stock concentration: 0.1–1.0 mg/mL.
• Mix gently, avoid vortexing.
• Aliquot and store at –20°C or –80°C.
• Dilute to working concentration in cell culture medium before use.
• Activate with APMA only if required for your assay.

These steps will ensure optimal solubility, stability, and activity of recombinant human MMP-1 for cell culture experiments.