Recombinant Human E3 ubiquitin-protein ligase ZNRF3 is produced through the baculovirus expression system and contains amino acids 56 to 219 of the protein. A C-terminal 6xHis-tag is attached to this partial protein, which makes purification and detection more straightforward. The product achieves purity levels above 90%, confirmed by SDS-PAGE analysis. This appears to provide high-quality material for research focused on understanding ubiquitin-mediated processes.
ZNRF3 functions as an E3 ubiquitin-protein ligase that's involved in regulating Wnt signaling pathways. The protein promotes ubiquitination and subsequent degradation of Frizzled receptors. This activity seems critical for modulating cellular signaling and maintaining tissue homeostasis. Studying this protein may be important for understanding how cells communicate and how signaling pathways contribute to various physiological processes.
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.
1. Protein-Protein Interaction Studies Using His-Tag Pull-Down Assays
The C-terminal 6xHis tag attached to this ZNRF3 fragment (56-219aa) allows for nickel-affinity purification and pull-down experiments to identify potential binding partners. This particular region might contain important protein interaction domains that could bind with Wnt signaling pathway components or other cellular proteins. With purity exceeding 90%, the fragment appears suitable for co-immunoprecipitation studies and in vitro binding assays. These experiments could potentially help clarify the molecular mechanisms behind ZNRF3's role in cellular signaling networks.
2. Antibody Development and Validation
This purified ZNRF3 fragment may serve as an immunogen for generating specific antibodies against the 56-219aa region of human ZNRF3. The high purity (>90%) and well-defined amino acid sequence suggest it's suitable for immunization protocols in research animals. The resulting antibodies could then be used for Western blotting, immunofluorescence, and other research applications to study endogenous ZNRF3 expression and localization patterns across different cell types and tissues.
3. Biochemical Characterization and Stability Studies
Researchers can use the recombinant ZNRF3 fragment for detailed biochemical analysis, including thermal stability, pH tolerance, and buffer compatibility studies. Such experiments would likely provide fundamental information about the protein's biophysical properties and optimal storage conditions. The His-tag makes purification relatively easy for multiple experimental conditions, allowing researchers to establish protocols for handling this protein domain in various research applications.
4. ELISA-Based Quantitative Assays
The His-tagged ZNRF3 fragment can be used to develop enzyme-linked immunosorbent assays (ELISA) for research purposes. The protein can be immobilized on ELISA plates either directly or through anti-His antibodies, which enables the development of sandwich or competitive assays. These assays might prove valuable for studying ZNRF3 interactions with other proteins or for validating antibody specificity in research settings.
