Recombinant Human Protein Wnt-2 (WNT2) is expressed in E.coli and covers the full length of the mature protein from amino acids 26 to 360. The protein carries an N-terminal 6xHis-tag, which aids in purification and detection. SDS-PAGE analysis confirms purity greater than 90%, suggesting reliable performance in research applications. This product is designed solely for research use and is not suitable for therapeutic or diagnostic purposes.
Wnt-2 belongs to the Wnt protein family, which appears to play a crucial role in regulating cellular processes like proliferation, differentiation, and migration. It participates in the canonical Wnt signaling pathway—a key pathway in developmental biology and cancer research. Understanding Wnt-2's function and interactions may be vital for exploring its potential implications in tissue regeneration and developmental disorders.
Potential Applications
Note: The applications listed below are based on what we know about this protein's biological functions, published research, and experience from experts in the field. However, we haven't fully tested all of these applications ourselves yet. We'd recommend running some preliminary tests first to make sure they work for your specific research goals.
Based on the provided information, the recombinant human WNT2 is expressed in E. coli, a prokaryotic system that is fundamentally unsuitable for producing functional eukaryotic signaling proteins like WNT2. WNT proteins require precise folding, multiple disulfide bond formations, and specific lipid modifications (palmitoylation) for their biological activity and receptor binding capability. E. coli lacks the eukaryotic chaperones, disulfide isomerases, and modification machinery necessary for proper WNT protein folding and processing. The WNT2 protein is expressed as the mature region (26-360aa) with an N-terminal 6xHis tag, but this represents only the polypeptide chain without essential post-translational modifications. The >90% purity indicates low contamination but does not confirm proper folding. Therefore, the folding status and functionality of this recombinant human WNT2 protein remain to be validated.
1. Antibody Development and Validation
This application is appropriate as the primary use case. The recombinant WNT2 can serve as an effective immunogen for generating antibodies that recognize linear epitopes within the 26-360aa region, even if the protein is misfolded. The His-tag facilitates purification and screening. However, antibodies may not recognize conformational or modification-dependent epitopes of properly folded, lipid-modified native WNT2. Validation against endogenous WNT2 from mammalian cells is essential.
2. Protein-Protein Interaction Studies
This application is highly problematic without folding validation. If this WNT2 protein is misfolded, it will not interact physiologically with its receptors (Frizzled, LRP5/6) or regulatory proteins. The multiple disulfide bonds required for proper WNT structure are unlikely to form correctly in E. coli. Identified interactions would likely be non-physiological artifacts. This application should not be pursued without confirmation of proper folding and receptor-binding activity.
3. Biochemical Characterization and Biophysical Analysis
This application is well-suited for assessing the recombinant human WNT2 protein itself. Techniques like circular dichroism spectroscopy, size-exclusion chromatography, and thermal shift assays can evaluate the protein's folding state and stability. However, these will characterize the E. coli-expressed product, which likely differs significantly from native, properly modified WNT2. These studies are valuable for quality control but cannot validate biological activity.
4. Cell-Based Binding and Uptake Studies
This application should be avoided without functional validation. If WNT2 is misfolded and unlipidated, it will not bind to Frizzled receptors or initiate signaling. Cell-based assays would yield false negative results and misleading conclusions about WNT2 function. This application requires prior demonstration of proper folding and receptor activation capability.
Final Recommendation & Action Plan
Given the extreme challenges of producing functional WNT proteins in E. coli, recommend the following approach: First, perform extensive biophysical characterization (circular dichroism for secondary structure, non-reducing SDS-PAGE for disulfide bond analysis) to assess folding quality. Antibody development can proceed as the safest application. Completely avoid all functional studies (interactions, cell-based assays) unless proper folding and biological activity are conclusively demonstrated through receptor-binding assays. For reliable WNT2 research, obtain the protein from mammalian or insect cell expression systems capable of proper disulfide bond formation and lipidation. Always include appropriate controls, such as active, properly modified WNT2 when possible.
