Cadherin 1, type 1, E-cadherin (epithelial), also known as CDH1 and CD32, is a tumor suppressor gene. 1, 2 The encoded protein is a calcium dependent cell-cell adhesion glycoprotein comprised of five extracellular cadherin repeats, a transmembrane region and a highly conserved cytoplasmic tail. E-cadherin-mediated cell-cell adhesion pathway may represent a novel chemotherapeutic target for bladder cancer, prostate cancer, and renal-cell carcinoma. 3
Protein Details
Purity
>95% by SDS Page and analyzed by silver stain.
Endotoxin Level
<0.1 EU/µg as determined by the LAL method
Biological Activity
The biological activity of Mouse E-CAD was determined by the ability of the immobilized protein to support the adhesion of the MCF7 human breast adenocarcinoma cells. When 5 x 10<sup>4</sup> cells/well are added to rmECAD coated plates (1.5 μg/ml with 100 μl/well), approximately 50-70% will adhere after 90 minutes at room temperature.
The predicted molecular weight of Recombinant Mouse ECAD is Mr 62 kDa. However, the actual molecular weight as observed by migration on SDS-PAGE is Mr 75-87 kDa.
Predicted Molecular Mass
62
Formulation
This recombinant protein was 0.2 µm filtered and lyophilized from a sterile solution containing 50mM Tris-Citrate, 50 mM NaCl and 2 mM CaCl2, pH 6.5.
Storage and Stability
This lyophilized protein is stable for six to twelve months when stored desiccated at -20°C to -70°C. After aseptic reconstitution, this protein may be stored at 2°C to 8°C for one month or at -20°C to -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 E-Cadherin is widely used in research applications because it is a critical mediator of calcium-dependent cell-cell adhesion in epithelial tissues, and its recombinant form enables precise experimental manipulation of cell signaling, adhesion, and differentiation processes.
Key scientific reasons to use recombinant mouse E-Cadherin include:
Modeling Cell-Cell Adhesion: E-Cadherin is essential for the formation and maintenance of adherens junctions in epithelial cells, which are fundamental for tissue integrity and barrier function. Recombinant protein allows controlled studies of these processes in vitro.
Stem Cell Research: E-Cadherin regulates pluripotency, self-renewal, and survival in mouse embryonic stem cells (mESCs). Manipulating E-Cadherin signaling with recombinant protein can direct stem cell differentiation, maintain pluripotency, or enhance survival in culture, which is valuable for regenerative medicine and developmental biology studies.
Differentiation Guidance: Recombinant E-Cadherin substratum can guide differentiation of embryonic stem cells into specific lineages, such as endoderm-derived hepatocyte-like cells, making it useful for cell fate studies and tissue engineering.
Cancer and EMT Studies: Loss or cleavage of E-Cadherin is associated with epithelial-to-mesenchymal transition (EMT) and cancer metastasis. Recombinant E-Cadherin enables functional assays to investigate these processes, including cell migration, invasion, and signaling pathway modulation.
Biophysical and Structural Analysis: Recombinant E-Cadherin is used in surface plasmon resonance, bioassays, and mechanistic studies to dissect its structural domains, binding partners (such as catenins), and signaling functions.
Experimental Control: Using recombinant protein provides a defined, reproducible system for studying E-Cadherin function, avoiding variability from endogenous expression or genetic background.
Typical applications include:
Coating culture surfaces to study cell adhesion and differentiation.
Supplementing cell culture media to modulate signaling pathways.
Functional assays for cell aggregation, migration, and barrier formation.
Biophysical studies of protein-protein interactions.
In summary, recombinant mouse E-Cadherin is a versatile tool for dissecting the molecular mechanisms of cell adhesion, stem cell biology, differentiation, and disease processes such as cancer and EMT, providing experimental control and reproducibility in a wide range of research contexts.
Yes, recombinant Mouse E-Cadherin can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately determined.
Supporting details:
ELISA kits for Mouse E-Cadherin routinely use recombinant protein as the standard. Multiple commercial ELISA kits specify lyophilized recombinant mouse E-Cadherin as the standard for generating calibration curves. This allows for quantitative measurement of E-Cadherin in biological samples by comparing sample signals to those from known concentrations of the recombinant standard.
Best practices for ELISA standards: It is recommended to use a purified protein for standard curve preparation. If a purified native protein is not available, a recombinant protein with verified purity and concentration is suitable. The recombinant standard should match the epitope recognized by the assay antibodies to ensure accurate quantification.
Validation and compatibility: ELISA kits are validated to recognize both natural and recombinant Mouse E-Cadherin, ensuring that recombinant standards are appropriate for calibration. The recombinant protein should ideally be produced in a system that yields a protein with similar post-translational modifications to the native protein, although most ELISA antibodies target linear epitopes, making this less critical for quantification.
Preparation: Follow the manufacturer’s instructions for reconstitution and dilution of the recombinant standard. Prepare a serial dilution to generate a standard curve covering the expected concentration range in your samples.
Carrier proteins: If using recombinant E-Cadherin as a standard, consider whether it contains carrier proteins (e.g., BSA), as these can affect stability and assay background. Carrier-free formulations are preferred when BSA or other additives may interfere with your assay.
Summary of protocol:
Reconstitute recombinant Mouse E-Cadherin according to the supplier’s instructions.
Prepare serial dilutions in the recommended buffer.
Run the standard curve alongside your samples in the ELISA.
Use the standard curve to interpolate sample concentrations.
In conclusion, recombinant Mouse E-Cadherin is scientifically appropriate and widely used as a standard for ELISA quantification, provided it is properly prepared and validated for your specific assay system.
Recombinant Mouse E-Cadherin has been validated in published research for several key applications, primarily in studies involving cell adhesion, stem cell biology, cancer research, and biophysical assays.
Validated Applications in Published Research:
Bioassays: Recombinant Mouse E-Cadherin has been widely used in cell-based bioassays to study cell-cell adhesion, epithelial barrier formation, and the mechanochemical regulation of cell behavior. Examples include:
Investigating self-assembly and morphogenesis in stem cell-derived synthetic embryos.
Studying mechanochemical control of epidermal stem cell divisions.
Assessing the effects of E-cadherin on cell division symmetry in stem cell colonies.
Evaluating the role of E-cadherin in combination therapies for diseases such as ulcerative colitis.
Western Blot Control: The protein is validated as a positive control in Western blotting to confirm the specificity of E-cadherin antibodies and to assess E-cadherin expression in mouse cell lysates.
Surface Plasmon Resonance (SPR): Recombinant Mouse E-Cadherin has been used in SPR assays to analyze protein-protein interactions, particularly to decipher structural and signaling functions of E-cadherin in mouse embryonic stem cells.
Cell Adhesion and Biophysical Studies: The protein is used in single-molecule force measurements (e.g., atomic force microscopy) to quantify E-cadherin-mediated adhesion and to study the effects of mutations or antibodies on E-cadherin binding strength.
ELISA Standard: It is recommended as a standard in ELISA for quantifying E-cadherin levels in biological samples, although direct literature citations for this use are less frequent compared to bioassays and Western blotting.
Additional Context:
Recombinant Mouse E-Cadherin is also used in studies of epithelial-to-mesenchymal transition (EMT), cancer metastasis, and tissue morphogenesis, reflecting its central role in cell-cell adhesion and signaling.
It serves as a tool for comparative immunology and preclinical therapeutic evaluations in mouse models.
Summary Table of Validated Applications
Application
Example Use Case/Publication Type
Bioassay
Stem cell morphogenesis, cell adhesion studies
Western Blot Control
Positive control for E-cadherin detection
Surface Plasmon Resonance
Protein-protein interaction analysis
Cell Adhesion/Biophysics
Single-molecule force measurements
ELISA Standard
Quantification of E-cadherin in samples
These applications are supported by multiple peer-reviewed studies and product validation summaries, confirming the versatility of recombinant mouse E-cadherin in both basic and translational research.
To reconstitute and prepare Recombinant Mouse E-Cadherin protein for cell culture experiments, add sterile water to the lyophilized powder to achieve the recommended stock concentration, typically 0.5 mg/mL or 250 μg/mL, depending on the specific formulation. Avoid vortexing; gently mix by pipetting or slow inversion to ensure complete dissolution.
Step-by-step protocol:
Reconstitution:
Add sterile water directly to the vial containing the lyophilized protein. For most preparations, a stock concentration of 0.5 mg/mL or 250 μg/mL is recommended.
Gently mix until fully dissolved. Do not vortex, as vigorous agitation can denature the protein.
If the formulation contains excipients (e.g., trehalose, mannitol, PBS, MES, NaCl, CaCl₂), these will help stabilize the protein during reconstitution.
Aliquoting and Storage:
Once reconstituted, aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
Store aliquots at –20°C to –80°C for long-term use. For short-term use (2–7 days), keep at 4–8°C.
If needed, add a carrier protein (e.g., 0.1% BSA) to further stabilize the solution, especially for low-concentration working stocks.
Preparation for Cell Culture:
Before use, dilute the stock solution in sterile, calcium-containing buffer or cell culture medium to the desired working concentration. E-Cadherin function is calcium-dependent, so ensure the buffer or medium contains sufficient Ca²⁺ (typically 1–2 mM).
For coating surfaces (e.g., adhesion assays), incubate the diluted protein on tissue culture plastic or glass at room temperature for 1–2 hours, then wash gently with buffer to remove unbound protein.
For direct addition to cell cultures, filter-sterilize the working solution if necessary and add to the medium under sterile conditions.
Additional notes:
Confirm the bioactivity of your preparation if functional assays are required, as some recombinant E-Cadherin proteins are not validated for activity.
Always consult the specific product datasheet or certificate of analysis for any manufacturer-recommended protocols or buffer requirements, as formulations may vary.
Summary of key points:
Reconstitute in sterile water to 0.5 mg/mL or 250 μg/mL.
Mix gently, do not vortex.
Aliquot and store at –20°C to –80°C; short-term at 4–8°C.
Dilute in calcium-containing buffer for cell culture use.
Avoid repeated freeze-thaw cycles.
These steps will ensure optimal solubility, stability, and functionality of recombinant mouse E-Cadherin for cell culture experiments.
References & Citations
1. Christofori, G. et al. (1998) Am. J. Hum. Genet. 63: 1588
2. Gumbiner, BM. et al. (2003) J. Cell Biol.161: 1191
3. Isaacs, WB. et al. (1995) World J Urol. 13:364
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
