Recombinant Human Cyclin-dependent kinases regulatory subunit 2 (CKS2) is expressed in E. coli, covering the full-length protein from amino acids 1 to 79. The protein carries an N-terminal 10xHis-tag and a C-terminal Myc-tag, which should help with purification and detection. SDS-PAGE analysis confirms the product shows purity greater than 90%, making it appropriate for various research applications.
CKS2 appears to be an integral component of the cell cycle. It plays what seems to be a crucial role in regulating cyclin-dependent kinases. Research suggests it's involved in cell cycle progression, and scientists have studied its interactions with other cell cycle proteins to better understand cell division mechanisms. This likely makes CKS2 a valuable protein for research in cellular regulation and cancer studies.
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 Pull-Down Assays
The dual-tagged CKS2 protein with N-terminal His-tag and C-terminal Myc-tag may enable comprehensive protein-protein interaction studies through pull-down experiments. The His-tag should allow for immobilization on nickel-affinity matrices to capture potential binding partners from cell lysates or purified protein libraries. Meanwhile, the Myc-tag provides an additional detection method for Western blot confirmation of successful protein capture and complex formation. This approach could help identify novel CKS2 interacting proteins or validate known interactions under controlled in vitro conditions.
2. Antibody Development and Validation
The recombinant CKS2 protein with >90% purity appears to serve as an excellent antigen for generating specific antibodies against human CKS2. Since it's the full-length protein (1-79aa), antibodies can likely be raised against various epitopes across the entire CKS2 sequence. The dual tags should allow for straightforward purification and quality control during antibody development processes. Generated antibodies can then be validated using the same recombinant protein in ELISA, Western blot, and immunoprecipitation assays to confirm specificity and binding affinity.
3. Biochemical Characterization and Stability Studies
The purified recombinant CKS2 protein enables detailed biochemical characterization. This might include thermal stability analysis, pH sensitivity studies, and buffer optimization experiments. Researchers can subject the protein to various analytical techniques such as dynamic light scattering, circular dichroism spectroscopy, and size exclusion chromatography to determine its biophysical properties. The dual-tag system appears to help with protein detection and quantification throughout these characterization studies, providing reliable methods for monitoring protein integrity and concentration.
4. In Vitro Binding Assays and Competition Studies
The tagged CKS2 protein can be used in plate-based binding assays to study its interaction with known binding partners or to screen for potential inhibitors. The His-tag should enable direct coating onto nickel-coated plates, while the Myc-tag allows for detection using anti-Myc antibodies in ELISA-format assays. Competition binding experiments may be performed to determine binding kinetics and to evaluate how various compounds or mutations affect CKS2 interactions.
