The specific activity, measured with 10 μM ES002 and 50 ng enzyme in 100 μL of TCNB (50 mM Tris, 10 mM CaCl2, 0.15 M NaCl, 0.05% Brij-35, pH 7.5) at room temperature, is > 300 pmol/min/μg.
The predicted molecular weight of Recombinant Mouse MMP-3 is Mr 52 kDa. However, the actual molecular weight as observed by migration on SDS-PAGE is Mr 60 kDa.
Predicted Molecular Mass
52
Formulation
Supplied frozen as a 0.2 μm filtered solution containing 25 mM Tris, 5 mM CaCl2, 50 mM NaCl, 0.02% Brij-35, 50% Glycerol, pH 8.0 at a concentration of 0.50 mg/ml.
Storage and Stability
This protein is stable for one month when stored sterile at -20°C or for three months when stored at -70°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles. See Product Insert for exact lot specific storage instructions.
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Recombinant Mouse MMP-3 is widely used in research applications due to its critical role in extracellular matrix (ECM) degradation, tissue remodeling, angiogenesis, and disease modeling. Employing the recombinant form ensures high purity, batch-to-batch consistency, and eliminates variability associated with tissue-derived enzymes.
Key scientific reasons to use recombinant Mouse MMP-3 include:
ECM Degradation and Tissue Remodeling: MMP-3 (Stromelysin-1) is a zinc-dependent endopeptidase that cleaves various ECM components such as collagen, fibronectin, and proteoglycans. This activity is essential for studying processes like wound healing, embryonic development, and immune responses.
Disease Modeling: MMP-3 is implicated in pathological conditions such as arthritis, cancer progression, atherosclerosis, and neurological diseases (e.g., stroke). Recombinant MMP-3 allows precise investigation of its role in these contexts, including its impact on inflammation, apoptosis, and vascular remodeling.
Mechanistic Studies: Recombinant MMP-3 is used to dissect molecular pathways, such as its activation of other MMPs (notably MMP-9), which is crucial for vascular smooth muscle cell migration and plaque stability in atherosclerosis models.
Cell Isolation and Tissue Dissociation: Purified recombinant MMPs, including MMP-3, are increasingly used as reagents for tissue dissociation, offering a defined alternative to crude collagenases of microbial origin.
Controlled Experimental Conditions: Recombinant proteins provide reproducible results in enzyme activity assays and mechanistic studies, supporting robust experimental design and interpretation.
Tagging and Detection: Recombinant MMP-3 can be engineered with tags (e.g., His-tag) for easy purification and detection, facilitating downstream applications such as protein-protein interaction studies and structural analysis.
In summary, using recombinant Mouse MMP-3 enables precise, reproducible, and mechanistic studies of ECM dynamics, disease processes, and cell biology, making it a valuable tool for basic and translational research.
Yes, recombinant Mouse MMP-3 can be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay system and properly validated. Recombinant proteins are commonly used as standards in ELISA kits for quantifying target analytes, including MMP-3.
Key considerations for use:
Protein Formulation: Recombinant Mouse MMP-3 is available in both carrier-free and carrier-added formulations. Carrier proteins (such as BSA) are often added to improve stability and handling. For ELISA standards, the formulation with BSA is generally recommended unless your assay is sensitive to carrier proteins.
Validation: Ensure the recombinant MMP-3 standard is validated for use in your specific ELISA format. Many commercial ELISA kits use recombinant Mouse MMP-3 as their calibration standard, indicating its suitability for this purpose.
Standard Curve Preparation: Prepare serial dilutions of the recombinant protein in the same buffer or matrix as your samples to generate a standard curve. This allows for accurate quantification of MMP-3 in unknown samples.
Activity and Isoform: Confirm whether your ELISA detects total MMP-3 (pro-, active, and complexed forms) or only specific isoforms. The recombinant standard should match the form(s) detected by your assay.
Unit Calibration: Recombinant MMP-3 is typically supplied as mass (ng or µg) and may also have activity units (pmol/min/µg). For ELISA quantification, use mass concentration for standard curve generation.
Best Practices:
Reconstitute and dilute the recombinant standard according to the ELISA kit instructions or your validated protocol.
Store aliquots of the recombinant protein at recommended conditions to maintain stability and avoid freeze-thaw cycles.
If using a custom ELISA, verify that the recombinant standard yields a linear and reproducible standard curve in your assay matrix.
Summary Table: Recombinant Mouse MMP-3 as ELISA Standard
Aspect
Recommendation/Note
Formulation
Use BSA-containing for ELISA unless interference occurs
Validation
Confirm compatibility with your ELISA system
Standard Curve
Prepare serial dilutions in assay buffer/matrix
Isoform
Match standard to ELISA detection specificity
Storage
Follow manufacturer’s instructions for stability
In conclusion, recombinant Mouse MMP-3 is widely used and suitable as a standard for ELISA quantification, provided it is matched to your assay’s requirements and validated for accuracy and reproducibility.
Recombinant Mouse MMP-3 has been validated for several applications in published research, primarily in functional bioassays, enzymatic activity studies, and mechanistic investigations of extracellular matrix remodeling and signaling.
Key validated applications include:
In vitro enzymatic assays: Recombinant Mouse MMP-3 is widely used to assess its proteolytic activity against various substrates, including extracellular matrix proteins such as collagen, fibronectin, laminin, aggrecan, and proteoglycans. These assays help characterize substrate specificity and catalytic efficiency.
Bioassays for substrate cleavage: It has been used to demonstrate dose-dependent degradation of specific proteins, such as nerve growth factor (NGF), in controlled in vitro settings. This validates its functional activity and allows quantification of its effects on target molecules.
Cell-based assays: Recombinant MMP-3 has been applied to cultured cells (e.g., human corneal endothelial cells) to study its effects on cell viability, extracellular matrix remodeling, and downstream signaling pathways. These studies often involve titration of recombinant protein to determine cytotoxicity thresholds and functional outcomes.
Tissue dissociation and cell isolation: Purified recombinant MMPs, including MMP-3, have been explored as reagents for tissue dissociation, offering an alternative to traditional collagenases for isolating cells from tissues.
ELISA standards and controls: Recombinant Mouse MMP-3 is used as a standard in ELISA assays to quantify endogenous MMP-3 levels in biological samples, and as a positive control in Western blotting and direct ELISA detection.
Mechanistic studies in disease models: In vivo and ex vivo studies have used recombinant or virally delivered MMP-3 to investigate its role in disease processes such as glaucoma (by modulating intraocular pressure and outflow facility), arthritis, wound healing, and cancer progression.
Bioassays for cytokine and enzyme regulation: MMP-3’s ability to activate other MMPs (e.g., pro-MMP-1, pro-MMP-7, pro-MMP-9, pro-MMP-13) and modulate cytokines (e.g., IL-1β, IGFBP-3) has been validated in biochemical assays.
Summary Table: Validated Applications of Recombinant Mouse MMP-3
Application Type
Example/Details
Enzymatic activity assays
Cleavage of ECM proteins, NGF, and other substrates
Cell-based functional assays
Effects on cell viability, ECM remodeling, signaling
Tissue dissociation
Cell isolation from tissues
ELISA/Western blot standards
Quantification and detection of MMP-3
Disease model mechanistic study
Modulation of IOP in glaucoma, arthritis, cancer, wound healing
Cytokine/enzyme regulation
Activation of other MMPs, cytokine processing
These applications are supported by peer-reviewed studies and product validation data, confirming the utility of recombinant Mouse MMP-3 in both basic and translational research contexts.
To reconstitute and prepare Recombinant Mouse MMP-3 protein for cell culture experiments, follow these steps:
Reconstitution
Briefly centrifuge the vial to collect the protein at the bottom before opening.
Reconstitute the lyophilized protein in 20 mM Tris, 150 mM NaCl, pH 8.0 to a final concentration of 0.1–1.0 mg/mL.
Do not vortex; gently mix by pipetting or slow inversion to avoid protein denaturation.
Activation (if required for your application)
MMP-3 is typically supplied as a pro-enzyme and may require activation for enzymatic activity.
To activate, dilute the reconstituted protein to 100 µg/mL in assay buffer containing 1 mM APMA (p-aminophenylmercuric acetate).
Incubate at 37 °C for 1 hour to achieve activation.
After activation, dilute to the desired working concentration in assay buffer or cell culture medium.
Buffer Compatibility for Cell Culture
For cell culture experiments, ensure the final buffer is compatible with your cells.
After activation, exchange the buffer to a cell culture-compatible medium (e.g., PBS or serum-free medium) using dialysis or a desalting column if necessary to remove APMA and excess salts.
Sterility
Filter the final protein solution through a 0.2 µm filter to ensure sterility before adding to cell cultures.
Storage
Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles.
Store at –80 °C for long-term storage. For short-term use, store at 4 °C.
Additional Notes:
Always confirm the endotoxin level is suitable for cell culture (<1 EU/µg is typical for sensitive applications).
Adjust the final concentration according to your experimental requirements; typical working concentrations for cell stimulation range from 1–100 ng/mL.
Avoid buffers containing surfactants (e.g., SDS) or organic solvents (e.g., methanol) when preparing protein for cell culture.
Summary of Key Steps:
Reconstitute in Tris/NaCl buffer, gently mix, activate with APMA if needed, exchange to cell-compatible buffer, filter for sterility, aliquot and store appropriately.
If your experiment requires the active form of MMP-3, ensure the activation step is performed. If using the protein for non-enzymatic purposes (e.g., as a standard in ELISA), activation may not be necessary. Always consult the specific datasheet for your recombinant protein batch for any manufacturer-specific recommendations.
References & Citations
1. Tang, CH. et al. (2011) J Cell Biochem. 112(5):1431-40.
2 Tang, CH. et al. (2012) J Cell Biochem. 113(6):1977-86.
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
