Recombinant Human Galectin-7

Recombinant human galectin-7 offers several compelling advantages for research applications across multiple biological disciplines.

Biological Significance and Functional Relevance

Galectin-7 is a prototype monomeric galectin with highly specialized expression patterns and regulatory functions that make it particularly valuable for studying epithelial biology and cellular homeostasis. This protein is specifically expressed in stratified epithelia, notably the epidermis, making it an ideal model for investigating epithelial-specific mechanisms.

The protein functions as a pro-apoptotic factor that operates intracellularly upstream of JNK activation and cytochrome-c release. This makes it essential for research into programmed cell death pathways and apoptotic regulation. Notably, galectin-7 increases the susceptibility of keratinocytes to UVB-induced apoptosis, a critical process in maintaining epidermal homeostasis.

Cell Adhesion and Migration Studies

Recombinant galectin-7 is particularly valuable for investigating cell-cell adhesion dynamics and collective cell behavior. The protein regulates E-cadherin endocytosis at the cell surface in a dose-dependent manner, controlling the turnover of this critical adhesion molecule. This regulatory function makes galectin-7 indispensable for studying epithelial cohesion and keratinocyte migration patterns.

Research demonstrates that galectin-7 depletion significantly impairs wound healing capacity and reduces cell migration directionality. By using recombinant galectin-7, you can perform rescue experiments to restore normal adhesion dynamics and quantitatively measure the protein's dose-dependent effects on cell behavior.

Extracellular Matrix Remodeling

Galectin-7 participates in extracellular matrix remodeling through interactions with matrix metalloproteinases, particularly MMP-2 and MMP-9. This makes the recombinant protein valuable for investigating tissue repair mechanisms, immune responses, and cancer cell migration associated with reduced cell adhesion.

Versatile Research Applications

The recombinant protein is suitable for multiple experimental approaches including SDS-PAGE analysis, bioassays, ELISA standards, and cell culture applications. Its availability in both carrier-free and carrier-protein formulations allows flexibility in experimental design depending on your specific requirements.

Genetic and Molecular Studies

Recombinant galectin-7 enables investigation of the LGALS7 gene and its regulation by tumor suppressor protein p53 following genotoxic events. The protein's high amino acid homology between human and mouse species (79%) facilitates translational research between in vitro and in vivo model systems.

Human galectin-7 is also relevant for studying immune modulation and barrier function, including its potential role in atopic dermatitis and skin barrier impairment. Additionally, the protein's involvement in innate and adaptive immune responses through glycan binding on immune cell surfaces expands its utility for immunological research.

Yes, recombinant human galectin-7 can be used as a standard for quantification and calibration in ELISA assays. This is one of the primary applications for recombinant galectin-7 proteins in research.

Formulation Considerations

When selecting recombinant galectin-7 for use as an ELISA standard, the formulation choice is critical. BSA-conjugated formulations are specifically recommended for use as ELISA standards, as they provide stability and compatibility with typical assay protocols. Conversely, carrier-free formulations should be reserved for applications where the presence of carrier proteins could interfere with your assay or downstream applications.

Standard Curve Development

To prepare your standard curve, you will typically start with a concentrated stock solution (commonly 10,000 pg/mL) and perform serial dilutions to establish your calibration range. The detection range for galectin-7 ELISA assays typically spans from approximately 125-10,000 pg/mL, depending on the specific kit configuration and detection method employed.

Quality Assurance

When using recombinant galectin-7 as your standard, ensure that your assay maintains appropriate precision and accuracy. Standard ELISA protocols require that internal controls be prepared in the same sample matrix as your experimental samples to account for potential matrix effects that could differ from the standard diluent alone. This validation step helps confirm that your standard curve accurately reflects the behavior of the analyte in your actual biological samples.

The sandwich ELISA principle used in galectin-7 assays is specifically designed to measure target protein concentrations by comparing sample optical density values against your prepared standard curve.

Research Applications and Validation

Recombinant human galectin-7 has been validated across multiple research applications, reflecting its diverse biological roles:

Cell Biology and Apoptosis Studies

The protein has been extensively used in bioassays to investigate apoptotic mechanisms. Recombinant galectin-7 demonstrates pro-apoptotic activity, functioning intracellularly upstream of JNK activation and cytochrome-c release. It has been validated for inducing apoptosis in T cell populations, with studies demonstrating that recombinant human galectin-7 effectively kills both Jurkat T cells and human peripheral T cells.

Cancer Research Applications

Recombinant galectin-7 has been applied in cancer immunotherapy research, particularly for investigating triple-negative breast cancer (TNBC) and other malignancies. The protein has been used to study galectin-7-induced T-cell apoptosis as a mechanism by which cancer cells evade immune surveillance. Additionally, research has validated its role in promoting tumorigenesis through upregulation of matrix metalloproteinase-9 (MMP-9) gene expression.

Wound Healing and Tissue Repair

The recombinant protein has been validated in studies examining epithelial wound repair mechanisms. Research has established that galectin-7 enhances endometrial re-epithelialization through integrin-mediated signaling and elevation of extracellular matrix factors.

Protein Interaction and Structural Studies

Recombinant galectin-7 has been used in protein-protein interaction studies and as a target for developing specific inhibitors, including nanobody-based therapeutics. These applications have validated its use in characterizing binding specificity and developing diagnostic imaging tools.

Standard Biochemical Assays

The protein is validated for use in SDS-PAGE analysis and serves as an ELISA standard. It has also been employed in agglutination assays measuring biological activity through red blood cell agglutination.

Reconstitution Guidelines

Recombinant Human Galectin-7 is typically supplied in lyophilized form and requires proper reconstitution before use in cell culture experiments. The standard reconstitution concentration is 200 μg/mL in sterile PBS. However, alternative reconstitution approaches are available depending on your specific experimental requirements.

For maximum flexibility, you can reconstitute the protein in deionized sterile water to achieve a stock concentration between 0.1–1.0 mg/mL. If you use this approach, you can subsequently dilute the stock solution in PBS (pH 7.4) to reach your desired working concentration. This method preserves the original salt concentration of the lyophilized formulation.

Formulation Considerations

The lyophilized protein is typically formulated with stabilizing agents including MOPS, NaCl, EDTA, DTT, and trehalose, with BSA as a carrier protein. Some formulations may include additional components such as Sarcosyl, proclin-300, or mannitol depending on the specific preparation.

Important handling notes:

• Do not vortex the reconstituted solution, as this can cause protein aggregation
• Centrifuge the vial prior to opening to collect all lyophilized material
• Avoid repeated freeze-thaw cycles, which compromise protein stability

Storage and Stability

After reconstitution, store the protein solution appropriately based on your timeline:

• Short-term storage: 4–8°C for 2–7 days
• Long-term storage: Aliquots at ≤ −20°C for up to 3 months
• Extended storage: −80°C for prolonged preservation

Use a manual defrost freezer and minimize freeze-thaw cycles to maintain protein integrity.

Enhancing Stability

Glycerol can be added to the reconstituted solution at a final concentration of 5–50% to improve protein stability and reduce flexibility of the N-terminal region. This is particularly useful if you plan extended storage or multiple handling steps in your cell culture experiments.

Preparation for Cell Culture Applications

When using the reconstituted protein in cell culture work, ensure the solution is sterile and free of particulates. If you are performing quantitative assays or immunological applications, verify that your reconstitution buffer is compatible with your downstream detection methods, as different buffers may affect protein-ligand interactions or antibody recognition.