Recombinant Chicken Homeobox protein engrailed-1 (EN1) is produced using an E.coli expression system and contains the full-length protein sequence from 1-333 amino acids. The protein carries an N-terminal 6xHis-B2M tag for purification and detection. SDS-PAGE analysis confirms greater than 90% purity, which makes it suitable for various research applications.
Homeobox protein engrailed-1 (EN1) appears to play a vital role in developmental processes and pattern formation. Research suggests it regulates gene expression during embryonic development, with particular effects on neural and skeletal structures. Given its critical function in these pathways, EN1 has become a significant focus of research in developmental biology and related fields.
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
This full-length recombinant chicken EN1 protein works well in pull-down assays to identify and characterize protein partners that interact with the engrailed-1 homeobox protein. The N-terminal 6xHis-B2M tag allows for immobilization on nickel-affinity resins, capturing interacting proteins from chicken embryonic tissue lysates or cell extracts. Such studies may help reveal the molecular mechanisms underlying EN1 function in developmental processes. The high purity (>90%) likely ensures reliable results with minimal background interference from contaminating proteins.
2. DNA-Protein Binding Assays
The recombinant EN1 protein serves as a useful tool for electrophoretic mobility shift assays (EMSA) or surface plasmon resonance studies to investigate its DNA-binding specificity and affinity. Researchers can test various DNA sequences containing potential engrailed binding sites to map the protein's recognition elements and binding kinetics. This application appears particularly valuable for studying transcriptional regulation mechanisms in avian development. The full-length construct (1-333aa) preserves the complete DNA-binding domain structure necessary for authentic binding activity.
3. Antibody Development and Validation
This purified recombinant protein can work as an immunogen for generating specific antibodies against chicken EN1 or as a positive control for validating existing anti-EN1 antibodies. The high purity level makes it suitable for immunization protocols in laboratory animals to produce polyclonal or monoclonal antibodies. The protein can also serve as a standard in Western blot, ELISA, or immunofluorescence experiments to confirm antibody specificity and determine optimal working concentrations. The His-B2M tag enables tag-specific detection methods for experimental validation as well.
4. Structural and Biophysical Characterization
Researchers can apply the recombinant EN1 protein in biophysical studies such as circular dichroism spectroscopy, dynamic light scattering, or analytical ultracentrifugation to characterize its folding state, secondary structure content, and oligomerization behavior. These analyses may provide insights into the protein's structural properties and stability under various buffer conditions. The E. coli expression system and high purity make it suitable for large-scale preparation needed for structural studies. Such characterization data would likely prove valuable for understanding structure-function relationships of this homeobox transcription factor.
5. In Vitro Transcription Assays
This full-length EN1 protein can be incorporated into cell-free transcription systems to study its role as a transcriptional regulator using chicken-specific promoter sequences. Researchers can examine how EN1 affects transcription initiation or elongation by adding the purified protein to reconstituted transcription reactions containing RNA polymerase II and associated factors. The complete amino acid sequence (1-333aa) ensures that both DNA-binding and transcriptional regulatory domains are present for functional studies. These assays may help define the mechanistic role of EN1 in gene expression control during avian development.
