Recombinant Human INO80D Protein: Research Guide & Applications
Recombinant Human INO80D Protein: Research Guide & Applications
Recombinant Human INO80D Protein is a key accessory subunit of the INO80 chromatin-remodeling complex, produced recombinantly for use in biochemical, structural, and functional studies of ATP-dependent chromatin regulation.
- Product Name
- Recombinant Human INO80D Protein
- SKU
- rp366675
- Protein Family
- INO80 chromatin-remodeling complex subunit
- Species Origin
- Human (Homo sapiens)
- Application
- Biochemical assays, protein–protein interaction studies, chromatin remodeling research
- Storage Recommendation
- Store at −80 °C; avoid repeated freeze-thaw cycles
- Form
- Recombinant protein (typical lyophilized or liquid format; confirm with CoA)
Biological Background of INO80D
INO80D (also referred to as TFPT or a designated subunit of the INO80 chromatin-remodeling complex) is a component of the evolutionarily conserved INO80 complex, an ATP-dependent chromatin remodeler belonging to the SNF2 family of proteins. The INO80 complex plays critical roles in DNA repair, replication, transcriptional regulation, and the maintenance of genomic stability by repositioning nucleosomes and facilitating access to DNA.
INO80D contributes to the structural integrity and regulatory specificity of the INO80 holocomplex. Research indicates that individual subunits of the INO80 complex, including INO80D, modulate the enzymatic activity of the catalytic ATPase subunit and determine substrate specificity. Understanding the precise function of INO80D is an active area of investigation in chromatin biology, with relevance to cancer biology, DNA damage response, and gene expression regulation.
Product Specifications and Quality
Aladdin Scientific's Recombinant Human INO80D Protein (SKU: rp366675) is produced using recombinant expression systems to ensure consistent batch-to-batch quality. Researchers should consult the Certificate of Analysis (CoA) supplied with each lot for precise details on:
- Purity: Assessed by SDS-PAGE and/or HPLC; typical recombinant proteins of this class achieve ≥85% purity (confirm with CoA).
- Biological Activity: Functional activity data, where available, is provided in the CoA.
- Formulation: Buffer composition, protein concentration, and format (lyophilized or liquid) are lot-specific; refer to the CoA.
- Endotoxin Level: Tested and reported per lot to ensure suitability for sensitive assays.
All specifications should be verified against the official CoA provided at time of shipment.
Handling, Storage, and Reconstitution Protocol
Proper handling of Recombinant Human INO80D Protein is essential to preserve its integrity and activity. Follow these general guidelines:
- Receipt: Upon receipt, briefly centrifuge the vial to collect contents. Inspect for any signs of damage or contamination.
- Storage: Store at −80 °C for long-term preservation. For short-term use (up to 1 week), storage at −20 °C may be acceptable. Avoid repeated freeze-thaw cycles, which can lead to protein aggregation and loss of activity.
- Reconstitution (lyophilized format): Reconstitute the lyophilized protein in sterile, ultrapure water or the recommended reconstitution buffer as specified in the CoA. Allow the vial to equilibrate to room temperature before opening. Add the reconstitution buffer slowly along the vial wall. Mix gently by rotating or pipetting; do not vortex.
- Aliquoting: After reconstitution, prepare single-use aliquots to minimize freeze-thaw cycles.
- Working Concentrations: Determine optimal working concentrations empirically for each assay. Refer to published literature for guidance on INO80 complex subunit assay conditions.
Note: Always consult the product-specific CoA and datasheet for lot-specific instructions that supersede these general guidelines.
Experimental Applications
Recombinant Human INO80D Protein is suitable for a range of research applications in chromatin biology and related fields:
- Protein–Protein Interaction Studies: Use in co-immunoprecipitation (Co-IP), pull-down assays, or surface plasmon resonance (SPR) to map interactions within the INO80 complex or with external binding partners.
- Biochemical Reconstitution: Incorporate INO80D into in vitro reconstituted INO80 complex assemblies to study subunit contributions to ATPase or nucleosome-remodeling activities.
- Structural Studies: Suitable as a component for cryo-EM or X-ray crystallography studies aimed at resolving the architecture of the INO80 complex.
- Antibody Validation: Use as a positive control antigen in Western blotting or ELISA to validate anti-INO80D antibodies.
- Drug Discovery: Screen small-molecule inhibitors or modulators targeting the INO80 complex using recombinant INO80D as a tool reagent.
Optimization of assay conditions is recommended for each application. Consult the relevant primary literature for experimental precedents.
Troubleshooting Common Issues
Researchers working with Recombinant Human INO80D Protein may encounter the following issues. The guidance below addresses common challenges:
| Issue | Possible Cause | Suggested Action |
|---|---|---|
| Protein aggregation | Repeated freeze-thaw cycles or improper storage | Prepare single-use aliquots; store at −80 °C |
| Low activity in functional assays | Protein degradation or suboptimal buffer conditions | Verify storage conditions; optimize buffer pH, salt concentration, and cofactors (e.g., ATP, Mg²⁺) |
| No signal in Western blot | Incorrect antibody, denaturation conditions, or insufficient protein loading | Use a validated anti-INO80D antibody; confirm protein loading with Coomassie or total protein stain |
| Inconsistent results between lots | Lot-to-lot variability | Always refer to the lot-specific CoA; run an internal positive control |
For unresolved technical issues, contact Aladdin Scientific technical support with your lot number and CoA details.
Frequently asked questions
What is the function of INO80D in the INO80 complex?
INO80D is an accessory subunit of the INO80 ATP-dependent chromatin-remodeling complex. It is thought to contribute to the structural assembly and regulatory specificity of the complex, modulating the ATPase activity of the catalytic subunit and influencing nucleosome-remodeling outcomes. Its precise molecular role remains an active area of research.
How should Recombinant Human INO80D Protein be stored to maintain activity?
Recombinant Human INO80D Protein should be stored at −80 °C for long-term stability. Avoid repeated freeze-thaw cycles by preparing single-use aliquots after reconstitution. Always refer to the Certificate of Analysis (CoA) for lot-specific storage and formulation instructions.
What applications is Recombinant Human INO80D Protein suitable for?
This protein is suitable for protein–protein interaction studies (Co-IP, pull-down, SPR), in vitro biochemical reconstitution of the INO80 complex, structural biology studies (cryo-EM, crystallography), antibody validation as a positive control antigen, and small-molecule screening in drug discovery programs targeting the INO80 complex.
How do I reconstitute lyophilized Recombinant Human INO80D Protein?
Allow the vial to equilibrate to room temperature before opening. Add the recommended reconstitution buffer (sterile ultrapure water or buffer as specified in the CoA) slowly along the vial wall. Mix gently by rotation or careful pipetting; do not vortex. Prepare single-use aliquots and store at −80 °C.
Where can I find purity and activity data for my specific lot of rp366675?
Purity, activity, formulation, and endotoxin data are provided in the Certificate of Analysis (CoA) specific to each lot. The CoA is included with shipment and may also be available by contacting Aladdin Scientific customer support with your lot number.
References
- Gerhold, C.B. & Gasser, S.M. INO80 and SWR complexes: relating structure to function in chromatin remodeling. Trends in Cell Biology, 2014.
- Watanabe, S. & Peterson, C.L. The INO80 family of chromatin-remodeling enzymes: regulators of histone variant dynamics. Cold Spring Harbor Symposia on Quantitative Biology, 2010.
- Aladdin Scientific. Recombinant Human INO80D Protein (rp366675) Certificate of Analysis. Aladdin Scientific, Shanghai.
Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.
危险品化学品经营许可证（带存储）