Recombinant Human Proliferation marker protein Ki-67 (MKI67) is produced in E. coli and includes an N-terminal 6xHis-tag for easier purification. This partial protein spans amino acids 2962-3254 and comes with purity levels above 90% as confirmed by SDS-PAGE. The product is intended for research use only and supports studies that need high-quality protein with minimal endotoxin content.
Ki-67 stands as a well-studied proliferation marker that researchers have examined extensively in cell biology. Understanding cellular processes like cell cycle progression appears to depend heavily on this protein, and scientists frequently turn to it for cell proliferation research. When cells actively divide, Ki-67 typically shows up, which likely explains why it has become such an important tool in cancer research and diagnostic work.
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.
Human Ki-67 is a large nuclear protein (358kDa) that functions in cell proliferation and requires complex folding and a specific nuclear environment for its biological activity. This recombinant fragment (2962-3254aa) represents only a small portion of the full protein. While E. coli can produce soluble proteins, the correct folding of this isolated domain cannot be guaranteed without experimental validation. Ki-67's function involves specific protein-protein interactions in the nucleolus, which this fragment may not replicate without the full protein context.
1. Antibody Development and Validation
This recombinant Ki-67 fragment serves as an excellent immunogen for generating antibodies specific to this C-terminal region (2962-3254aa) of human Ki-67. The high purity and defined sequence ensure targeted antibody production against linear epitopes. These antibodies will be particularly useful for Western blot applications to detect proteolytic fragments or specific domains of Ki-67 in research samples.
2. Biochemical Characterization and Biophysical Analysis
This is the essential first step to assess the protein's physical properties. Techniques like size-exclusion chromatography can determine oligomeric state and homogeneity, while circular dichroism can analyze secondary structure content and thermal stability. These studies provide critical quality control data for understanding this specific fragment's behavior, not the native protein.
3. ELISA Development and Immunoassay Applications
This protein is well-suited as a standard for quantitative ELISA assays targeting this specific Ki-67 region. The assay depends on antibody binding to linear epitopes, so the protein's folding state is less critical. The His-tag facilitates consistent immobilization for reliable standard curve generation in research immunoassays.
Final Recommendation & Action Plan
This recombinant Ki-67 fragment is valuable for immunological and analytical applications but should be used with caution for interaction studies due to uncertainties about its folding state. The recommended approach is to prioritize Application 2 (Biochemical Characterization) to assess the protein's physical properties and folding quality. Applications 1 and 3 (Antibody Development and ELISA) can proceed confidently as they rely on sequence-specific recognition rather than native structure. For protein-protein interaction studies, preliminary validation of folding and interaction capability with a known binding partner is recommended before large-scale screening. This protein is primarily suitable as an antigen source and immunoassay standard rather than for functional studies requiring native Ki-67 activity.
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