Matrix metallopeptidase 2, also known as MMP2, is an enzyme involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes, such as arthritis and metastasis. Most MMP's are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. MMP-2 degrades type IV collagen, the major structural component of basement membranes and plays a role in endometrial menstrual breakdown, regulation of vascularization and the inflammatory response. MMP2 has been found to be associated with tumor invasion in various cancers.1
Protein Details
Purity
>95% by SDS-PAGE and analyzed by silver stain.
Endotoxin Level
<1.0 EU/µg as determined by the LAL method
Biological Activity
The specific activity, measured under the described conditions, is > 1,500 pmoles/min/μg.
The predicted molecular weight of Recombinant Mouse MMP-2 is Mr 71 kDa. However, the actual molecular weight as observed by migration on SDS-PAGE is Mr 72 kDa.
Predicted Molecular Mass
71
Formulation
Supplied as a 0.2 μm filtered solution of 25 mM Tris, pH 7.5, 5 mM CaCl2, 75 mM NaCl, 0.025% Brij-35 and 50% glycerol at a concentration of 0.113 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.
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Recombinant Mouse/Rat MMP-2 is widely used in research applications because it enables precise investigation of the biological roles of MMP-2 in extracellular matrix (ECM) remodeling, inflammation, axonal regeneration, angiogenesis, and cancer progression, using controlled and reproducible experimental conditions.
Key scientific applications and rationale include:
Axonal Regeneration and Neurobiology: Recombinant MMP-2 facilitates the study of axonal regrowth mechanisms, particularly in models of CNS injury. It degrades inhibitory ECM components such as chondroitin sulfate proteoglycans (CSPGs) in glial scars, thereby promoting axonal regeneration. Administration of recombinant MMP-2 to dissociated adult rat retinal ganglion cells (RGCs) has been shown to stimulate axonal regrowth, making it a valuable tool for dissecting molecular pathways underlying neural repair.
ECM Remodeling: MMP-2 is a type IV collagenase that degrades collagens, elastin, and fibronectin, which are major components of the ECM. Recombinant MMP-2 allows researchers to model and quantify ECM degradation in vitro, facilitating studies on cell migration, tissue remodeling, and wound healing.
Cancer Biology: MMP-2 is critical for tumor invasion, metastasis, and angiogenesis by breaking down basement membranes and ECM barriers. Recombinant MMP-2 is used to investigate these processes, screen inhibitors, and study the modulation of the tumor microenvironment, which is essential for developing anti-metastatic therapies and understanding cancer progression.
Inflammation and Immune Response: MMP-2 modulates inflammatory responses by regulating immune cell migration and phenotype. Recombinant protein enables mechanistic studies on how MMP-2 influences immune cell behavior and inflammatory gene expression, relevant for cardiovascular, autoimmune, and neuroinflammatory diseases.
Controlled Experimental Design: Using recombinant MMP-2 ensures batch-to-batch consistency, defined activity, and absence of confounding factors present in tissue extracts. This is crucial for reproducibility and for dissecting specific biochemical pathways in cell-based and biochemical assays.
Developmental Biology: MMP-2 is involved in developmental processes such as axonal outgrowth and navigation. Recombinant protein allows for precise manipulation of MMP-2 activity in developmental models, aiding in the study of morphogenesis and tissue patterning.
In summary, recombinant Mouse/Rat MMP-2 is essential for mechanistic studies in ECM biology, neuroregeneration, cancer research, and inflammation, providing a reliable and defined tool for probing MMP-2 function in diverse experimental systems.
Yes, recombinant Mouse/Rat MMP-2 protein can be used as a standard for quantification or calibration in ELISA assays designed to measure MMP-2. This approach is widely accepted in research applications for both mouse and rat samples.
Supporting details:
ELISA kits for MMP-2 quantification commonly use recombinant mouse/rat MMP-2 as the calibration standard. For example, the Quantikine Total MMP-2 Immunoassay is calibrated against a highly purified recombinant mouse/rat pro-MMP-2, and the kit documentation confirms accurate quantitation of both recombinant and natural MMP-2.
Standard curves generated with recombinant MMP-2 are shown to be parallel to those obtained with natural MMP-2, indicating that the recombinant protein is suitable for determining relative mass values of endogenous MMP-2 in biological samples.
Recovery and linearity data from these kits demonstrate that recombinant MMP-2 spiked into various sample matrices (cell culture media, serum, plasma) is recovered at rates close to 100%, supporting its use for calibration.
Best practices: Ensure the recombinant standard matches the species and isoform of MMP-2 you wish to quantify, and confirm that your ELISA antibodies recognize both recombinant and native forms. Most commercial kits are validated for this purpose, but custom assays should be similarly validated.
Additional considerations:
Use recombinant MMP-2 expressed in a system that preserves relevant post-translational modifications if your assay is sensitive to these features.
For research use only; recombinant standards are not suitable for clinical diagnostics unless specifically validated for such use.
Summary: Recombinant Mouse/Rat MMP-2 is a scientifically validated standard for ELISA quantification and calibration, provided the assay is designed to recognize both recombinant and native forms of the protein.
Recombinant Mouse/Rat MMP-2 has been validated for several key applications in published research, primarily in studies of enzyme activity, protein detection, and functional assays related to extracellular matrix remodeling and disease models.
Validated Applications in Published Research:
Enzyme Activity Assays: Recombinant Mouse/Rat MMP-2 is widely used as a positive control or standard in gelatin zymography and other enzyme activity assays to measure gelatinase activity, particularly for distinguishing pro- and active forms of MMP-2 in biological samples. These assays are critical for studying MMP-2’s role in tissue remodeling, vascular biology, and disease processes.
Western Blotting: The recombinant protein is used to validate antibody specificity and as a molecular weight marker for MMP-2 in Western blot analyses. For example, it has been used to confirm the detection of both pro- and active forms of MMP-2 in muscle tissue and to validate the performance of anti-MMP-2 antibodies.
ELISA (Enzyme-Linked Immunosorbent Assay): Recombinant MMP-2 serves as a standard or positive control in direct ELISAs for quantifying MMP-2 levels in biological samples.
Functional and Mechanistic Studies: The protein is used in mechanistic studies to investigate the role of MMP-2 in physiological and pathological processes, such as:
Retinal angiogenesis and neovascularization: Recombinant MMP-2 has been used in rodent models to study its involvement in retinal neovascularization and as a target for pharmacological inhibition.
Vascular remodeling: Used in studies examining the effects of mechanical stretch and reactive oxygen species on MMP-2 expression and activity in vascular smooth muscle cells.
Myogenesis: Research has shown MMP-2 is necessary for myogenic gene activation and myoblast differentiation, implicating its use in studies of muscle development and regeneration.
Summary Table of Validated Applications
Application
Description/Context
Enzyme Activity Assays
Gelatin zymography, activity quantification, positive control
Western Blotting
Antibody validation, detection of pro/active MMP-2, molecular weight marker
ELISA
Standard/positive control for quantification assays
Recombinant Mouse/Rat MMP-2 is often used interchangeably in mouse and rat studies due to their high sequence homology (99%).
It is typically supplied at high purity (>95%) and validated for biological activity in these applications.
In published research, it is rarely used for in vivo administration but is essential for in vitro and ex vivo biochemical and cell-based assays.
If you require protocols or more specific details for any of these applications, please specify the context or experimental system.
To reconstitute and prepare Recombinant Mouse/Rat MMP-2 protein for cell culture experiments, follow these steps:
Reconstitution: Briefly centrifuge the vial to ensure all lyophilized protein is at the bottom. Add sterile distilled water (ddH₂O) to achieve a final concentration of 0.1–1.0 mg/mL. Do not vortex; gently mix by pipetting or slow inversion to avoid protein denaturation.
Incubation: Allow the protein to dissolve completely at room temperature for 10–15 minutes, mixing gently to ensure full reconstitution.
Activation (if required): MMP-2 is often supplied as a proenzyme (inactive zymogen). To activate, dilute the reconstituted protein to 40 μg/mL in assay buffer containing 1 mM APMA (4-aminophenylmercuric acetate) and incubate at 37°C for 1–2 hours. This step converts pro-MMP-2 to its active form, which is necessary for functional assays.
Buffer Exchange (optional): If the protein is supplied in a buffer containing glycerol, detergents, or other additives, consider buffer exchange (e.g., via dialysis or spin columns) into a cell culture-compatible buffer (such as PBS or serum-free medium) to avoid cytotoxicity.
Sterility: Filter the final solution through a 0.2 μm sterile filter if sterility is required for cell culture applications.
Aliquoting and Storage: Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles. Store aliquots at –80°C for long-term storage or 2–8°C for short-term use (up to one week). Avoid repeated freeze/thaw cycles to maintain protein integrity.
Working Concentration: Dilute the activated MMP-2 to the desired working concentration in cell culture medium immediately before use. Typical concentrations for cell-based assays range from 10–100 ng/mL, but optimal dosing should be determined empirically for your specific experiment.
Always confirm the specific formulation and recommended reconstitution protocol from the product datasheet, as buffer composition and concentration may vary by supplier.
If using for cell culture, ensure all reagents and buffers are sterile and endotoxin-free.
For functional assays, activation with APMA is essential unless the protein is supplied in active form.
If you need a protocol for a specific cell type or assay, please specify for tailored recommendations.
References & Citations
1. Viswanadha, VP. et al. (2017) J Cell Biochem. 118(9):2865-2876.
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
