Recombinant Human Angiopoietin-related protein 6 (ANGPTL6) is produced in E. coli and includes the complete sequence of the mature protein spanning amino acids 21 to 470. This recombinant protein carries a C-terminal 6xHis-tag, which makes purification and detection more straightforward. The product shows purity greater than 90%, confirmed through SDS-PAGE analysis. This level of purity appears suitable for demanding research applications.
Angiopoietin-related protein 6 (ANGPTL6) belongs to the angiopoietin-like family, which seems to influence various physiological processes. Research suggests ANGPTL6 may be involved in regulating metabolism and angiogenesis. Scientists studying metabolic pathways and angiogenesis-related processes are likely to find this protein particularly useful for their investigations.
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, the folding state and bioactivity of this recombinant ANGPTL6 protein are unknown and must be considered highly uncertain. ANGPTL6 is a secreted glycoprotein that typically undergoes complex folding, including the formation of oligomers and specific disulfide bonds essential for its activity. Expression in the reducing cytoplasm of E. coli makes correct disulfide bond formation unlikely, and the absence of eukaryotic glycosylation further diverges from its native post-translational modification pattern. While the protein represents the mature sequence (21-470aa), the C-terminal His-tag could potentially interfere with the structure of the C-terminal fibrinogen-like domain, which is critical for function. Therefore, it is highly improbable that this protein is correctly folded and bioactive. Applications relying on specific biological interactions or activities are speculative without validation.
1. Antibody Development and Validation
This recombinant ANGPTL6 protein is suitable for use as an immunogen to generate antibodies. Antibodies generated will be against the linear epitopes of this non-glycosylated, misfolded protein. Their ability to recognize the correctly folded, glycosylated, and oligomeric native ANGPTL6 in human serum or tissues is very low and must be rigorously tested. The protein is reliable as a positive control for Western blotting (denaturing conditions) but not for detecting the native protein.
2. Biochemical Characterization and Stability Studies
This purified recombinant protein is well-suited for basic biochemical and biophysical characterization. Researchers can determine its oligomerization state, thermal stability, and aggregation behavior under various buffer conditions. This application is valid because it focuses on the intrinsic physical properties of the purified polypeptide, which are independent of its native bioactivity.
3. ELISA-Based Quantitative Assays
The His-tagged ANGPTL6 can be used to develop an ELISA. An ELISA built with this recombinant human ANGPTL6 protein would only detect itself or antibodies raised against it. It would not accurately quantify native ANGPTL6, leading to erroneous results. Its use in ELISA should be restricted to testing antibodies specific to the immunogen.
Final Recommendation & Action Plan
The recommended course of action is to restrict the use of this protein lot to applications that do not depend on native conformation. It can be used for antibody production (Application 1, with the explicit caveat that the antibodies may not recognize the native protein) and for biophysical characterization of the recombinant material itself (Application 2). Protein-protein interaction studies rely on the protein having native-like structure and function, which are not feasible and should be abandoned. The essential first step to enable functional studies is to obtain a version of the protein produced in a mammalian expression system, which can facilitate proper folding, oligomerization, and glycosylation. Before any quantitative or interaction studies are attempted, the activity of any new protein preparation must be validated in a functional assay, such as one measuring its effect on endothelial cell migration or proliferation.
