Recombinant Human Nuclear cap-binding protein subunit 1 (NCBP1) is expressed in a baculovirus system, covering the full length of 1-790 amino acids. The protein carries an N-terminal 10xHis-tag and a C-terminal Myc-tag, which streamline purification and detection in experimental applications. SDS-PAGE analysis indicates the product achieves greater than 85% purity, making it suitable for various research applications that demand high-quality reagents.
NCBP1 appears to be an essential component of the cap-binding complex and plays a critical role in mRNA processing and stability. The protein is involved in the initial steps of mRNA transport from the nucleus to the cytoplasm and is vital for efficient mRNA splicing and translation. NCBP1's function in these pathways makes it an important protein for studies on gene expression regulation and cellular response mechanisms.
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. RNA Cap-Binding Complex Assembly and Characterization Studies
This full-length NCBP1 protein can be used to reconstitute the nuclear cap-binding complex (CBC) in vitro by co-expressing or mixing with NCBP2 (CBP20). The dual His and Myc tags enable purification and detection in pull-down assays to study protein-protein interactions within the CBC. Researchers may investigate the stoichiometry, binding kinetics, and structural requirements for CBC formation using biochemical approaches. The baculovirus expression system typically provides proper eukaryotic folding, which likely makes this protein suitable for functional complex assembly studies.
2. mRNA Cap Structure Binding Assays
The recombinant NCBP1 can be used in in vitro binding assays to study interactions with 7-methylguanosine cap structures on mRNA. Cap-analog resins or synthetic capped RNA substrates allow researchers to characterize binding specificity and affinity. The N-terminal His tag streamlines protein immobilization for surface plasmon resonance or other biophysical binding studies. These assays can help elucidate the molecular basis of cap recognition and the role of different NCBP1 domains in RNA binding.
3. Antibody Development and Validation
The dual-tagged full-length NCBP1 serves as an excellent antigen for generating specific antibodies against human NCBP1. Its high purity (>85%) and full-length nature ensure comprehensive epitope coverage for polyclonal antibody production. Both Myc and His tags can be used as positive controls in immunoassays to validate antibody specificity and cross-reactivity. Generated antibodies can subsequently be used for immunoprecipitation, Western blotting, and immunofluorescence studies of endogenous NCBP1 in cell biology research.
4. Protein-Protein Interaction Screening
The tagged NCBP1 protein can be used in pull-down assays to identify novel interacting partners involved in mRNA processing and nuclear export pathways. The His tag enables immobilization on metal affinity resins for capturing potential binding partners from cell lysates or purified protein libraries. Meanwhile, the Myc tag allows for easy detection and quantification of the bait protein in these interaction studies. This approach may help map the NCBP1 interactome and discover new components of cap-dependent RNA metabolism pathways.
5. Structural and Biophysical Characterization Studies
This full-length recombinant NCBP1 provides material for detailed structural studies including X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy analysis. The protein can be used to investigate conformational changes upon RNA binding or complex formation with NCBP2. Biophysical techniques such as dynamic light scattering, analytical ultracentrifugation, and thermal stability assays can characterize the protein's oligomerization state and stability. The baculovirus expression system's post-translational modification capabilities may preserve native-like protein structure for these analyses.
