Recombinant Human P-Cadherin

Using Recombinant Human P-Cadherin in research applications is valuable for studying cell adhesion, tissue architecture, cancer biology, and therapeutic targeting due to its defined molecular properties and biological relevance.

Key scientific reasons to use recombinant human P-cadherin:

• Cell Adhesion and Tissue Architecture: P-cadherin is a classical cadherin that mediates calcium-dependent cell-cell adhesion, playing a crucial role in maintaining tissue structure by linking to the actin cytoskeleton through catenins. Recombinant forms allow controlled studies of these interactions in vitro.

• Cancer Research: Overexpression of P-cadherin is associated with tumor aggressiveness, enhanced cell migration, invasion, and poor prognosis in several cancers, including breast and ovarian cancer. Recombinant P-cadherin enables mechanistic studies on how its expression influences tumor progression, metastasis, and cell signaling.

• Therapeutic Targeting and Drug Development: Recombinant P-cadherin is used to screen and characterize antibodies or small molecules that disrupt its adhesive function, which is a promising strategy for inhibiting cancer cell growth and metastasis. It also serves as a target antigen in the development and validation of bispecific antibodies and other biologics.

• Functional Assays: Recombinant P-cadherin supports cell adhesion assays, migration studies, and bioassays to dissect its role in cell signaling and collective cell migration. It provides a standardized, reproducible reagent for quantitative and comparative analyses.

• Mechanistic Studies: Using recombinant protein allows for precise manipulation of experimental conditions, such as mutagenesis or domain swapping, to dissect the molecular mechanisms of P-cadherin-mediated adhesion and signaling.

• Quality and Consistency: Recombinant proteins offer batch-to-batch consistency, defined sequence, and reduced variability compared to native or hybridoma-derived materials, which is critical for reproducible research outcomes.

Common applications include:

• Cell adhesion and migration assays
• Cancer cell invasion and metastasis models
• Antibody and inhibitor screening
• Structural and biophysical studies of cadherin interactions
• Mechanistic studies of cell-cell junctions and cytoskeletal dynamics

In summary, recombinant human P-cadherin is a versatile and essential tool for dissecting the molecular and cellular functions of P-cadherin in both normal physiology and disease contexts, especially cancer.

Yes, recombinant Human P-Cadherin can be used as a standard for quantification or calibration in ELISA assays, provided it is validated for this purpose and matches the assay’s requirements. Recombinant P-Cadherin is commonly supplied as the reference standard in commercial ELISA kits for quantitative measurement of P-Cadherin in biological samples.

Key considerations for use as an ELISA standard:

• Validation: The recombinant protein should be validated for use in ELISA, ensuring it is recognized by the capture and detection antibodies used in your assay. Most commercial ELISA kits use recombinant Human P-Cadherin as the standard, confirming its suitability for calibration.
• Purity and Form: The protein should be of high purity (typically >80% by SDS-PAGE) and in a form compatible with your assay (e.g., glycosylation status, tag presence). Some recombinant standards may have tags (e.g., His-tag) or be Fc chimeras; ensure these do not interfere with antibody recognition.
• Concentration and Dilution: Accurately reconstitute and dilute the recombinant standard according to your assay’s protocol to generate a reliable standard curve. The concentration range should cover the expected sample concentrations.
• Matrix Effects: If your samples are in complex matrices (serum, plasma, cell culture supernatant), confirm that the recombinant standard behaves similarly to endogenous P-Cadherin in those matrices. Recovery and linearity data from kit manuals often show that recombinant P-Cadherin is suitable for spiking and quantification in these sample types.
• Biological Activity: For quantification, biological activity is not required; only immunoreactivity with the assay antibodies is necessary.

Protocol best practices:

• Prepare the standard according to the manufacturer’s instructions or validated lab protocols, ensuring complete dissolution and proper mixing.
• Include multiple standard concentrations to generate a robust calibration curve, typically spanning the assay’s detection range (e.g., 24.7–18,000 pg/mL).
• Run standards in duplicate or triplicate to ensure precision and reproducibility.

Limitations:

• If using a recombinant standard not specifically validated for your ELISA kit, perform preliminary experiments to confirm its suitability, as differences in protein folding, glycosylation, or tags may affect antibody recognition.
• Always check for potential interference or cross-reactivity, although most kits report high specificity for Human P-Cadherin.

Summary Table: Recombinant Human P-Cadherin as ELISA Standard

In conclusion, recombinant Human P-Cadherin is widely used and suitable as a standard for ELISA quantification, provided it is validated for your specific assay system and protocol.

Recombinant Human P-Cadherin has been validated for several key applications in published research, primarily in the context of cancer biology, cell adhesion, and epithelial morphogenesis.

Validated Applications:

• Cell Adhesion Assays: Recombinant P-Cadherin is widely used to study cell-cell adhesion mechanisms, particularly in cancer models where P-Cadherin overexpression correlates with tumor aggressiveness and poor prognosis.
• Bioassays: Functional assays involving recombinant P-Cadherin have been employed to investigate its role in cell signaling, migration, and invasion, especially in breast cancer and other epithelial tumors.
• ELISA (Enzyme-Linked Immunosorbent Assay): Recombinant P-Cadherin serves as a standard or antigen in ELISA protocols for quantifying protein levels in biological samples.
• Western Blot (WB): Used as a positive control or standard for protein detection and quantification in Western blotting.
• Immunohistochemistry (IHC) and Immunofluorescence (IF/ICC): While these applications are more commonly validated for antibodies, recombinant P-Cadherin is sometimes used for antigen retrieval or as a control in tissue staining protocols.
• In Vivo Imaging and Tumor Targeting: Engineered forms of recombinant P-Cadherin have been used in molecular imaging studies to assess biodistribution, tumor targeting, and pharmacodynamics in animal models.
• Epithelial Morphogenesis Studies: Recombinant P-Cadherin has been utilized to investigate its essential role in lumen formation and cell protrusion during epithelial tissue development.

Supporting Details:

• Cancer Research: P-Cadherin is frequently overexpressed in high-grade carcinomas, and recombinant forms are used to dissect its contribution to tumor cell migration, invasion, and metastasis.
• Therapeutic Target Validation: Recombinant P-Cadherin is instrumental in validating antibody-based therapies that disrupt cell adhesion and suppress malignant growth.
• Developmental Biology: Studies have shown that P-Cadherin-dependent adhesions are required for proper epithelial morphogenesis, with recombinant protein used to model these processes in vitro.

Summary Table of Applications

Additional Notes:

• Recombinant P-Cadherin is not typically used for diagnostic or therapeutic purposes in clinical settings, but is a valuable tool for mechanistic and translational research.
• Its use in cell culture applications is well established, especially for dissecting adhesion-dependent signaling pathways.

If you require protocols or specific experimental details for any of these applications, please specify the context or assay type.

To reconstitute and prepare Recombinant Human P-Cadherin protein for cell culture experiments, gently resuspend the lyophilized protein in sterile distilled water or PBS to a concentration between 0.1–0.5 mg/mL, avoiding vigorous mixing or vortexing. After reconstitution, allow the protein to dissolve at room temperature with gentle agitation for 15–30 minutes.

Detailed protocol and best practices:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Add sterile distilled water or PBS (check the product datasheet for preferred buffer; PBS pH 7.4 is commonly used) to achieve the desired concentration (typically 0.1–0.5 mg/mL for cell culture applications).
• Gently mix by slow pipetting or gentle inversion. Do not vortex or pipette vigorously, as this may denature the protein.
• Let the solution sit at room temperature for 15–30 minutes to ensure complete dissolution.
• Inspect for clarity: If the solution is not clear, gently rock or invert until fully dissolved. Avoid foaming.
• Sterile filtration (0.22 μm) may be performed if required for cell culture, but only after complete dissolution.
• Aliquot and store the reconstituted protein at 2–8°C for short-term use (up to 1 week), or at –20°C to –80°C for longer-term storage. Avoid repeated freeze-thaw cycles.
• Working concentration: For coating plates or functional assays, typical concentrations range from 1–10 μg/mL, but optimal amounts depend on cell type and experimental design.

Additional notes for cell culture experiments:

• If using for substrate coating (e.g., adhesion assays), dilute the reconstituted protein in PBS or appropriate buffer to the desired working concentration, apply to culture surfaces, and incubate as per assay protocol.
• For soluble treatments, dilute the protein in cell culture medium immediately before use.
• Always use sterile technique to prevent contamination.

Summary of key steps:

• Centrifuge vial → Add sterile water/PBS → Gentle mixing → Room temp incubation → Aliquot/store appropriately → Dilute for experimental use.

If your specific application requires a different buffer or concentration, consult the product datasheet or relevant literature for optimal conditions.