Recombinant Human Friend leukemia integration 1 transcription factor (FLI1) is expressed in E. coli and contains amino acids 1-452, representing the full-length protein. The protein includes an N-terminal 10xHis tag to aid in purification, with a purity level above 85% as determined by SDS-PAGE. This product is for research use only and should not be used in diagnostic or therapeutic applications.
FLI1 is a transcription factor that regulates gene expression and appears to play a crucial role in blood cell development and vascular tissue formation. It belongs to the ETS family of transcription factors, which bind to specific DNA sequences and influence various cellular pathways. FLI1 has gained attention in research primarily because of its involvement in cellular differentiation and proliferation.
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. Protein-DNA Interaction Studies
This full-length recombinant human FLI1 protein can help researchers investigate its DNA-binding properties and sequence specificity using electrophoretic mobility shift assays (EMSA) and surface plasmon resonance studies. The N-terminal His-tag makes protein purification and immobilization more straightforward for these binding experiments. Scientists can examine how FLI1 interacts with different DNA sequences, including ETS-binding sites and potential regulatory elements. Such studies may contribute to our understanding of FLI1's role as a transcription factor in normal blood cell development and cancer-related processes.
2. Antibody Development and Validation
The purified recombinant FLI1 protein works well as an antigen for generating specific antibodies against human FLI1 through standard immunization protocols. The high purity level (>85%) helps reduce contamination that might lead to cross-reactive antibodies. Researchers can use the His-tagged protein in ELISA-based screening to identify and characterize antibody specificity and binding strength. These antibodies would likely prove valuable for immunoprecipitation, Western blotting, and immunofluorescence studies that investigate FLI1 expression patterns and cellular location.
3. Protein-Protein Interaction Analysis
This recombinant FLI1 protein can be used in pull-down assays to identify and study protein partners that interact with FLI1 in transcriptional complexes. The N-terminal His-tag allows for immobilization on nickel-affinity matrices, which can capture interacting proteins from cell extracts or purified protein libraries. Co-immunoprecipitation studies and yeast two-hybrid screens can be validated using this purified protein as a positive control. These experiments might help reveal the molecular mechanisms behind FLI1's transcriptional regulatory functions.
4. Structural and Biophysical Characterization
The full-length recombinant FLI1 protein provides material for structural biology studies, including X-ray crystallography, NMR spectroscopy, and cryo-electron microscopy experiments. Biophysical techniques such as dynamic light scattering, analytical ultracentrifugation, and thermal stability assays can characterize protein folding, oligomerization states, and stability. Having sufficient quantities of purified protein makes systematic structure-function relationship studies possible through limited proteolysis and domain mapping experiments.
5. In Vitro Transcription Assays
This recombinant FLI1 protein can be added to cell-free transcription systems to study its transcriptional regulatory activity on target promoters. The protein can be introduced to nuclear extracts or reconstituted transcription systems containing RNA polymerase II and associated factors to assess its impact on gene expression. Chromatin immunoprecipitation-sequencing (ChIP-seq) validation experiments can use this protein as a standard for testing antibody specificity. These assays may provide insights into FLI1's direct transcriptional targets and regulatory mechanisms.
