Recombinant Human Desmoglein-2

Recombinant Human Desmoglein-2 (DSG2) is a valuable tool in research due to its critical roles in cell adhesion, signaling, and disease modeling, particularly in studies of epithelial biology, cardiomyocyte function, cancer, and viral infection.

Key reasons to use Recombinant Human Desmoglein-2 in research applications:

• Cell Adhesion and Junction Studies: DSG2 is a major component of desmosomes, mediating calcium-dependent cell–cell adhesion in epithelial and cardiac tissues. Recombinant DSG2 allows for in vitro assays to study cell adhesion, cohesion, and the molecular mechanisms underlying tissue integrity.

• Cardiac Research: DSG2 is essential for cardiomyocyte cohesion and function. Recombinant DSG2 can be used in dissociation assays to assess the effects of mutations or drugs on cardiac cell cohesion, which is relevant for understanding arrhythmogenic cardiomyopathies.

• Cancer Biology: Altered DSG2 expression or function is implicated in tumor progression, invasion, and metastasis. Recombinant DSG2 enables mechanistic studies of cell–cell adhesion loss, epithelial-to-mesenchymal transition (EMT), and vasculogenic mimicry in cancer models.

• Viral Receptor Studies: DSG2 serves as a high-affinity receptor for certain adenovirus serotypes (e.g., Ad3, Ad7, Ad11, Ad14). Recombinant DSG2 is used to characterize virus–host interactions, screen for viral inhibitors, and model infection mechanisms.

• Signaling Pathways: DSG2 modulates intracellular signaling, including the Rap1 and TGFβ2 pathways, affecting cell spreading, migration, and focal adhesion protein phosphorylation. Recombinant DSG2 is useful for dissecting these pathways in cell-based assays.

• Assay Development: Recombinant DSG2 is suitable as a standard or capture protein in ELISA, Western blot, and cell adhesion assays, supporting quantitative and qualitative analyses of DSG2 interactions and function.

• Drug Discovery and Screening: It can be used to screen for molecules that modulate DSG2-mediated adhesion or signaling, which is relevant for therapeutic development in cancer, cardiac, and infectious diseases.

• Functional Validation: Recombinant DSG2 enables loss- and gain-of-function studies, helping to validate the role of DSG2 in various biological processes and disease models.

In summary, using recombinant human DSG2 provides a controlled, reproducible reagent for dissecting the molecular and cellular functions of desmoglein-2 in health and disease, facilitating a wide range of experimental applications in cell biology, virology, oncology, and cardiovascular research.

Yes, you can use Recombinant Human Desmoglein-2 as a standard for quantification or calibration in your ELISA assays, provided that the recombinant protein is highly purified and its concentration can be accurately determined. According to best practices for ELISA standard curve preparation, a purified protein—such as a recombinant protein—is suitable for generating a reliable standard curve, as long as it is quantified using a method like HPLC or spectrophotometry to ensure accuracy.

However, there are a few important considerations:

• Purity and Identity: The recombinant protein should be free of contaminants and should match the epitope(s) recognized by your ELISA detection antibodies.
• Concentration Determination: The concentration of the recombinant standard must be measured precisely, as inaccuracies here will propagate into your sample quantification.
• Matrix Effects: If possible, reconstitute and dilute the recombinant standard in a matrix similar to your samples (e.g., assay diluent or buffer) to minimize matrix interference.
• Validation: It is recommended to validate the performance of your recombinant standard by comparing it to a commercially available, certified standard if available.

In summary, Recombinant Human Desmoglein-2 is appropriate for use as an ELISA standard, as long as it is purified, accurately quantified, and compatible with your assay system.

Recombinant Human Desmoglein-2 (DSG2) has been validated for several applications in published research, primarily focusing on its biological and functional roles in cell adhesion, signaling, and as a viral receptor. Key applications supported by published studies include:

1. Cell Adhesion Assays:
   Recombinant DSG2 has been used to study cell-to-cell adhesion, particularly in epithelial and endothelial cells. For example, immobilized recombinant DSG2 supports the adhesion of human skin fibroblast cells (BUD8), demonstrating its functional activity in mediating cellular adhesion (Leinco, Cusabio).

2. Viral Receptor Studies:
   Recombinant DSG2 is used to investigate adenovirus infection mechanisms. It has been shown to act as a high-affinity receptor for adenovirus serotypes 3, 7, 11, and 14. Recombinant DSG2 protein can block adenovirus binding and infection, confirming its role as a functional viral receptor (Wang et al., 2010; Feng et al., 2020; PMC3074512, PMC7431812).

3. Signaling Pathway Analysis:
   Recombinant DSG2 is employed to study intracellular signaling pathways, such as the Dsg2-Rap1-TGFβ axis, which regulates cell spreading and focal adhesion dynamics. These studies often involve examining phosphorylation events in focal adhesion proteins like Paxillin and FAK (Shelton et al., 2021; Scientific Reports, 2021).

4. Barrier Function Studies:
   Recombinant DSG2 is used in research on epithelial barrier integrity, particularly in intestinal and cardiac tissues. It helps elucidate the role of DSG2 in maintaining cell cohesion and barrier function (Ungewiß et al., 2017; Schlegel et al., 2010; Cardiovascular Research, 2014).

5. Immunological and Antibody Validation:
   Recombinant DSG2 is used as a target for antibody validation and immunological assays, including immunofluorescence and Western blotting, to study DSG2 expression and localization (Nature Scientific Reports, 2017; R&D Systems, Bio-Techne).

6. Cancer and Vasculogenic Mimicry Research:
   Recombinant DSG2 is applied in studies exploring its role in cancer progression, particularly in melanoma, where it promotes vasculogenic mimicry and is associated with poor clinical outcomes (Tan et al., 2016; Oncotarget).

7. Structural and Functional Studies:
   Recombinant DSG2 is used in atomic-force microscopy and other biophysical assays to study the mechanics of DSG2-mediated adhesion and its interaction with other proteins (Schlegel et al., 2010; MOCA RWTH Aachen).

These applications highlight the versatility of recombinant human Desmoglein-2 in both basic research and translational studies, particularly in cell biology, virology, and cancer research.

To reconstitute and prepare Recombinant Human Desmoglein-2 (DSG2) protein for cell culture experiments, follow these best-practice steps:

1. Reconstitution

• Briefly centrifuge the vial before opening to ensure all lyophilized material is at the bottom.
• Add sterile deionized or distilled water (ddH₂O) to achieve a final concentration of 0.1–1.0 mg/mL.
• Do not vortex; instead, gently pipette up and down or swirl to dissolve the protein completely.
• Allow the protein to fully dissolve at room temperature, which may take 15–30 minutes. If undissolved material remains, gentle mixing for up to 2 hours at room temperature is acceptable.

2. Buffer Considerations

• If your application requires a specific buffer (e.g., PBS for cell culture), you may reconstitute directly in sterile PBS or first dissolve in water and then dialyze or dilute into the desired buffer.
• For long-term stability or to prevent aggregation, adding 5–50% glycerol can be beneficial, especially if storing at −20°C or below.

3. Sterility

• Use only sterile reagents and equipment throughout the process to maintain protein and cell culture sterility.
• If sterility is critical, filter the reconstituted solution through a 0.22 μm sterile filter before use.

4. Storage

• Aliquot the reconstituted protein to avoid repeated freeze/thaw cycles.
• Store at 2–8°C for up to one month, or at −20°C to −80°C for longer-term storage.
• Avoid repeated freeze/thaw cycles, as these can degrade the protein.

5. Application in Cell Culture

• For coating plates (e.g., for adhesion assays), a typical working concentration is 10 μg/mL in PBS.
• Incubate coated plates at 4°C overnight or at 37°C for 1–2 hours, then wash with PBS before adding cells.
• For soluble applications, dilute the reconstituted protein to the desired working concentration in cell culture medium immediately before use.

Summary Table: Key Steps

Notes:

• Always consult the specific product datasheet for any unique instructions.
• If the protein is tagged (e.g., His-tag), ensure compatibility with your downstream application.

These steps will help ensure optimal solubility, stability, and biological activity of recombinant DSG2 for cell culture experiments.