Recombinant Human Histone H2A.x (H2AFX) is produced in E. coli and includes the complete protein sequence spanning amino acids 1 to 143. The protein carries an N-terminal GST tag that helps with purification and detection processes. SDS-PAGE analysis confirms the product maintains purity levels above 90%, which appears to support reliable research outcomes. This protein is designed strictly for research purposes and should not be used in diagnostic or therapeutic applications.
Histone H2A.x represents a variant within the histone H2A family and seems to play an essential role in how cells respond to and repair DNA damage. When DNA sustains damage, H2A.x undergoes phosphorylation and likely helps recruit repair proteins to the damaged sites. This involvement in protecting genomic integrity makes it particularly valuable for research into cell cycle control and cancer studies.
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. Chromatin Remodeling and Nucleosome Assembly Studies
Researchers can use this full-length recombinant H2AFX protein in laboratory settings to examine chromatin dynamics and nucleosome formation processes. The protein may be incorporated into nucleosome reconstitution experiments along with other core histones (H2A, H2B, H3, H4) and DNA to study how chromatin structures develop. Scientists might investigate whether H2AFX behaves differently from standard H2A regarding nucleosome stability and chromatin packaging. The GST tag simplifies purification and handling during these intricate biochemical procedures.
2. GST Pull-Down Assays for Protein-Protein Interaction Studies
The N-terminal GST tag makes this protein immediately useful for GST pull-down experiments aimed at identifying and characterizing proteins that interact with H2AFX. Cell lysates or purified candidate proteins can be mixed with GST-H2AFX attached to glutathione beads to capture specific binding partners. This method proves especially valuable when studying histone chaperones, chromatin remodeling complexes, or DNA repair proteins that might bind to H2AFX. The greater than 90% purity should minimize background binding from contaminating proteins.
3. Antibody Development and Validation
This recombinant protein works well as an immunogen for creating H2AFX-specific antibodies or for testing existing antibodies. Since it contains the full-length sequence (1-143aa), it provides all potential epitopes found in the natural protein, making it suitable for producing antibodies that recognize different regions of H2AFX. Researchers can also use the protein in ELISA-based tests to assess antibody specificity, binding strength, and potential cross-reactivity with other histone variants.
4. In Vitro Histone Modification Assays
The recombinant H2AFX protein may serve as a substrate for investigating post-translational modifications, especially phosphorylation events connected to DNA damage response pathways. Kinase experiments can be conducted using this protein to study enzyme specificity and modification rates. Scientists might also use the protein to test the activity of histone-modifying enzymes like acetyltransferases, methyltransferases, or deacetylases that target H2AFX, potentially revealing new insights into epigenetic regulation mechanisms.
