Recombinant Human Histone H2AX (H2AFX) is produced in E. coli and includes the complete protein sequence from amino acids 1 to 143. The protein carries an N-terminal 6xHis-GST tag that helps with purification, reaching purity levels above 85% as confirmed by SDS-PAGE. This product is designed for research purposes only and appears to offer a dependable choice for different experimental setups.
Histone H2AX represents a variant within the histone H2A protein family and seems to play an essential role in how cells respond to DNA damage. It participates in DNA repair processes, especially when DNA double-strand breaks occur. When damage happens, H2AX gets phosphorylated, which likely marks the damage site and brings in repair proteins. Studying this protein may be critical for grasping how genomic stability works and how cells react to DNA damage.
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. In Vitro Chromatin Assembly and Nucleosome Reconstitution Studies
This full-length recombinant H2AX protein works well for reconstituting nucleosomes in the lab by mixing it with other core histones (H2A, H2B, H3, and H4) plus DNA substrates. The N-terminal His-GST tag makes purification and handling straightforward during chromatin assembly procedures. Scientists can examine how efficiently H2AX gets incorporated into nucleosome structures. They might also compare chromatin behavior between H2AX-containing and standard H2A-containing nucleosomes. These experiments could provide basic insights into chromatin organization and help clarify H2AX's structural role in chromatin fiber formation.
2. Protein-Protein Interaction Studies Using Pull-Down Assays
The N-terminal His-GST tag appears ideal for pull-down experiments aimed at finding and characterizing proteins that bind to H2AX. Scientists can attach the GST tag to glutathione-sepharose beads, while the His tag offers another purification and attachment option through nickel-based resins. Nuclear extracts or purified candidate proteins can help map the H2AX interactome under different conditions. This method seems particularly useful for studying how various cellular situations or post-translational modifications might affect H2AX protein partnerships.
3. Antibody Development and Validation
This recombinant H2AX protein makes an excellent antigen for creating specific antibodies against human H2AX or for testing existing ones. The high purity (>85%) and full-length design suggest that antibodies will recognize natural epitopes found in cellular H2AX. The dual tagging system lets researchers tell the difference between antibodies that recognize H2AX itself and those that might react with the tags instead. This application appears essential for developing trustworthy research tools in chromatin biology studies.
4. Biochemical Characterization and Biophysical Analysis
The purified recombinant protein works for detailed biochemical and biophysical studies of H2AX properties. This includes thermal stability, protein folding analysis, and structural examination. Scientists can apply techniques like circular dichroism spectroscopy, dynamic light scattering, or analytical ultracentrifugation to understand how H2AX behaves in solution. The His-GST tag makes protein handling and concentration measurements easier, while the high purity should ensure reliable and consistent experimental results. Such studies may help explain the fundamental properties that set H2AX apart from standard H2A histones.
5. In Vitro Enzyme Substrate Studies
This recombinant H2AX can work as a substrate for studying enzymes that modify histone proteins—kinases, phosphatases, ubiquitin ligases, or other chromatin-modifying enzymes. The purified protein creates a controlled system for investigating enzyme specificity, reaction rates, and optimal conditions without dealing with the complexity of cellular chromatin. The tags make detection and purification of modified products simpler for downstream analysis through mass spectrometry or other analytical methods. This application seems valuable for characterizing the enzymatic machinery involved in H2AX regulation and chromatin remodeling processes.
