Recombinant Mouse Isthmin-1 (Ism1) gets expressed in a yeast system and covers the full length of the mature protein, spanning amino acids 30 to 454. The protein comes with an N-terminal 6xHis tag, which makes purification and detection much easier. Purity levels reach above 90%, confirmed through SDS-PAGE analysis. This appears to ensure reliable performance for research work. The product is prepared under strict quality standards, which likely makes it appropriate for different experimental conditions.
Isthmin-1 (Ism1) has drawn attention in biological research because it seems to be involved in cellular processes like cell adhesion and angiogenesis. The protein may play an important role in regulating these pathways, helping scientists understand tissue development and repair mechanisms better. Researchers often work with Ism1 to explore how it functions and what it interacts with. This provides valuable insights into its significance in both normal physiology and disease states.
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.
Based on the provided information, recombinant mouse Ism1 is produced in a yeast expression system as the full-length mature protein (30-454aa) with an N-terminal 6xHis-tag. Yeast expression systems provide eukaryotic folding machinery capable of supporting disulfide bond formation and some post-translational modifications, which are important for Ism1's structural integrity and function. However, Ism1 is a complex secreted protein with multiple functional domains that require precise folding for its biological activities in cell signaling and development. The N-terminal His-tag may potentially interfere with the protein's native structure or function. No validation data (e.g., cell-based activity assays, receptor binding tests) are provided. Therefore, while yeast expression increases the probability of correct folding compared to prokaryotic systems, the protein's folding status and bioactivity cannot be confirmed without experimental validation.
1. Protein-Protein Interaction Studies
If the recombinant Ism1 is correctly folded, the His-tag enables pull-down assays to identify binding partners, as proper folding is essential for biologically relevant interactions. However, if misfolded, interaction domains may be altered, leading to non-specific binding or failure to recognize genuine biological partners, compromising the validity of identified networks. The high purity reduces background, but cannot compensate for structural defects.
2. Antibody Development and Validation
This application is suitable as antibody generation primarily relies on linear epitope recognition, which is independent of folding status. The full-length mature protein provides comprehensive epitope coverage for producing specific antibodies. However, if misfolded, generated antibodies may not optimally recognize conformation-dependent epitopes of native Ism1 in biological contexts.
3. Structural and Biochemical Characterization
If properly folded, the recombinant Ism1 is suitable for biophysical studies to understand its structural properties and stability. However, if misfolded, data from techniques like circular dichroism or dynamic light scattering would misrepresent the native protein's architecture, leading to incorrect conclusions about Ism1's structure-function relationships.
4. Cell-Based Functional Assays
If correctly folded and bioactive, the recombinant Ism1 can be used in cell culture systems to study its effects on cellular processes. However, if misfolded or inactive, it will not elicit proper biological responses, potentially leading to false-negative results or misinterpretation of Ism1's role in signaling pathways.
Final Recommendation & Action Plan
Before employing this recombinant Ism1 in any application, it is essential to validate its folding and bioactivity through biophysical methods (e.g., circular dichroism for secondary structure, size-exclusion chromatography for oligomeric state) and functional assays (e.g., testing known Ism1 activities in appropriate cell models); if validation confirms proper folding and function, proceed with applications while noting that the N-terminal His-tag may still affect some biological interactions; if misfolded or inactive, consider using alternative expression systems (e.g., mammalian cells) that may better support proper folding and post-translational modifications, or obtain a commercially validated standard; for immediate use, antibody development can proceed with the understanding that antibodies may require additional validation against native protein. Always include appropriate controls in experiments to ensure reliability.
