Recombinant Human Apoptosis regulator BAX (BAX) comes from an E.coli-based cell-free expression system, which produces a complete protein spanning amino acids 1-192. The protein carries a 10xHis-tag at its N-terminal end, making purification and detection more straightforward. SDS-PAGE analysis shows purity levels above 90%, suggesting this product should work well for research purposes. The protein is made using a detergent platform and includes a transmembrane domain.
BAX appears to be a key player in controlling apoptosis - it's a pro-apoptotic member of the Bcl-2 protein family. This protein seems particularly important for mitochondrial membrane permeabilization, where it helps release cytochrome c and trigger caspase activation. These steps are essential for programmed cell death. Given its role in apoptosis pathways, BAX likely holds significance for cancer research, neurodegenerative disease studies, and understanding how cells respond to stress.
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 recombinant Human BAX is expressed using an in vitro E. coli expression system (cell-free system), which can promote proper protein folding due to the absence of cellular complexities and chaperones. The protein is full-length (1-192aa), including all functional domains, and has an N-terminal 10xHis tag. The purity is >90% by SDS-PAGE. However, since activity is unverified, the protein cannot be assumed to be correctly folded or bioactive without functional validation. BAX requires precise folding for its oligomerization and membrane insertion functions in apoptosis. While cell-free systems often yield well-folded proteins, experimental confirmation is essential to ensure bioactivity (e.g., via oligomerization assays or membrane binding tests).
1. Membrane Protein Reconstitution Studies
The full-length BAX protein contains transmembrane domains and can be integrated into artificial lipid bilayers for reconstitution studies. However, if BAX is misfolded, its membrane insertion behavior may not reflect physiological conditions, leading to inaccurate kinetics or stability data. The His-tag facilitates purification and tracking, but folding must be validated first to ensure native-like behavior. If correctly folded, this application is feasible; otherwise, results may be misleading.
2. Protein-Protein Interaction Screening
The N-terminal 10xHis tag enables pull-down assays to identify binding partners (e.g., other BCL-2 family proteins). The high purity (>90%) reduces background noise. However, if BAX is misfolded, interactions may be non-physiological, yielding false positives or negatives. This application should be pursued only after confirming folding and activity, as BAX's interactions are conformation-dependent.
3. Antibody Development and Validation
This application is well-supported. The recombinant BAX can serve as an immunogen for antibody generation, even if misfolded, as antibodies may recognize linear epitopes. The full-length sequence ensures coverage of multiple antigenic regions. However, antibodies produced may not bind native BAX if conformational epitopes are altered, so validation against endogenous BAX is recommended. The His-tag simplifies purification for immunization.
4. Membrane Insertion Kinetics Analysis
This application is highly dependent on correct folding. If BAX is properly folded, fluorescence-based techniques can study insertion kinetics into membranes. However, if misfolded, kinetic data (e.g., insertion rates or structural changes) may not reflect biological reality, leading to erroneous conclusions. The His-tag aids concentration measurement, but functional validation is a prerequisite for reliable results.
Final Recommendation & Action Plan
Given the uncertainty in folding and bioactivity, the priority is to validate the protein's conformation and function using biophysical and biochemical assays. Recommend performing size-exclusion chromatography (to check oligomeric state), circular dichroism (for secondary structure), and functional tests (e.g., membrane binding or oligomerization assays) to confirm activity. If validated, the protein can be used for all described applications; if not, focus on antibody development and avoid membrane-related studies. The cell-free expression system is advantageous for folding, but verification remains critical. Always include controls such as known BAX activators or inhibitors in experiments.
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