Recombinant Mouse RNA polymerase II subunit A C-terminal domain phosphatase (Ctdp1) is expressed in E. coli and covers the amino acid region 178-341. This partial protein comes engineered with an N-terminal 10xHis tag and a C-terminal Myc tag, which makes purification and detection more straightforward. SDS-PAGE analysis confirms the product shows purity greater than 90%, suggesting high-quality performance for research applications.
Ctdp1 appears to be a crucial phosphatase involved in regulating RNA polymerase II. It plays what seems to be a significant role in transcriptional processes. The protein is known for dephosphorylating the C-terminal domain of RNA polymerase II, which likely influences transcription elongation and termination. Understanding this protein may be essential for transcriptional regulation studies and their broader implications in gene expression research.
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 Phosphatase Activity Assays
This recombinant Ctdp1 fragment (178-341aa) can help investigate phosphatase activity against RNA polymerase II C-terminal domain (CTD) substrates under controlled in vitro conditions. The dual His and Myc tags make purification and detection easier, which should streamline enzyme kinetics studies with various phosphorylated CTD peptides or proteins. Researchers might examine substrate specificity, optimal reaction conditions, and screen for inhibitors using this purified enzyme preparation. The high purity (>90%) appears to support reliable and reproducible biochemical assays for understanding Ctdp1 enzymatic properties.
2. Protein-Protein Interaction Studies
The dual-tagged Ctdp1 fragment works well as bait or prey in pull-down assays to identify novel protein interactions within the RNA polymerase II transcriptional machinery. The N-terminal His tag allows for immobilization on nickel-based resins. Meanwhile, the C-terminal Myc tag enables detection and validation of binding partners through immunoblotting. This approach may help map the protein interaction network of Ctdp1 and identify co-factors or regulatory proteins that modulate its function in transcriptional regulation.
3. Antibody Development and Validation
This purified recombinant protein fragment works as an immunogen for generating specific antibodies against mouse Ctdp1. It can also serve as a standard for validating existing antibodies. The defined amino acid region (178-341aa) provides a specific epitope source for antibody production targeting this domain of Ctdp1. High purity and dual tags support quality control assays, including ELISA-based antibody characterization and specificity testing through immunoblotting applications.
4. Structural and Biophysical Characterization
The purified Ctdp1 fragment can be used in structural biology approaches such as X-ray crystallography, NMR spectroscopy, or cryo-electron microscopy to understand the molecular architecture of this phosphatase domain. Biophysical techniques including dynamic light scattering, analytical ultracentrifugation, and thermal stability assays may provide insights into protein folding, oligomerization state, and stability. The dual tags help with protein tracking during purification and concentration steps required for structural studies.
