Recombinant Human FRYL Protein: Research Guide & Applications
Recombinant Human FRYL Protein: Research Guide & Applications
Recombinant Human FRYL Protein is a research-grade recombinant protein derived from the human FRYL gene, designed to support studies in chromatin organization, epigenetic regulation, and developmental signaling pathways.
- Product Name
- Recombinant Human FRYL Protein
- SKU
- rp373177
- Gene Name
- FRYL (FRY-like transcription coactivator)
- Species Origin
- Human (Homo sapiens)
- Application Area
- Epigenetics, chromatin regulation, developmental biology research
- Product Form
- Recombinant protein (specific formulation details: see product datasheet)
Background: The FRYL Gene and Protein
FRYL (FRY-like transcription coactivator) is a large scaffold protein encoded by the FRYL gene in humans. It is a paralog of FRY (Furry), a conserved protein originally characterized in Drosophila melanogaster as essential for maintaining cellular morphology and regulating the NDR (nuclear Dbf2-related) family of serine/threonine kinases. In mammals, FRYL plays roles in transcriptional coactivation, cell polarity, and developmental processes.
FRYL has been implicated in the regulation of the Hippo signaling pathway and interacts with NDR kinases (LATS1/2 and NDR1/2) to influence downstream targets including YAP and TAZ, which are key effectors of tissue growth and organ size control. Emerging research also connects FRYL to chromatin remodeling and epigenetic regulatory complexes, making recombinant FRYL protein a valuable tool for biochemical and cell biology studies.
Protein Specifications and Quality Considerations
Aladdin Scientific's Recombinant Human FRYL Protein (SKU: rp373177) is produced using recombinant expression systems to ensure consistent batch-to-batch quality. Researchers should consult the product datasheet for precise specifications including:
- Expression system: Refer to the product datasheet for host cell information (e.g., E. coli, baculovirus/insect cell, or mammalian expression system).
- Purity: Confirmed by SDS-PAGE and/or HPLC analysis; specific purity values are stated on the Certificate of Analysis (CoA).
- Formulation and storage buffer: See datasheet for buffer composition, stabilizers, and recommended storage temperature.
- Molecular weight: FRYL is a large protein (predicted >200 kDa based on the full-length gene); actual observed molecular weight may differ based on construct design; verify on CoA.
Always verify lot-specific details by reviewing the CoA provided with each shipment.
Recommended Handling and Storage Protocol
Proper handling of recombinant proteins is critical to preserve activity and integrity. Follow these general best practices for Recombinant Human FRYL Protein:
- Upon receipt: Briefly centrifuge the vial before opening to collect all material at the bottom. Inspect for any signs of damage or improper storage during transit.
- Reconstitution: If lyophilized, reconstitute according to the datasheet instructions using the recommended volume of the specified buffer. Allow the protein to dissolve gently; avoid vigorous vortexing.
- Aliquoting: Prepare single-use aliquots to prevent repeated freeze-thaw cycles, which can lead to protein aggregation and loss of activity.
- Storage: Store at –80 °C for long-term preservation. Short-term storage (≤1 week) may be acceptable at –20 °C; refer to the datasheet for product-specific recommendations.
- Freeze-thaw cycles: Minimize freeze-thaw cycles to ≤3 to maintain protein integrity. Addition of carrier protein (e.g., 0.1% BSA) may be recommended for highly dilute working solutions.
- Working conditions: Keep protein on ice during experimental setup. Avoid prolonged exposure to room temperature.
Research Applications
Recombinant Human FRYL Protein is suitable for a range of in vitro and biochemical research applications, including but not limited to:
- Protein–protein interaction studies: FRYL's scaffold function makes it a candidate for co-immunoprecipitation (Co-IP), pull-down assays, and surface plasmon resonance (SPR) experiments to map interaction partners within the NDR kinase and Hippo signaling networks.
- Biochemical activity assays: Use as substrate or regulatory component in kinase assays involving LATS1/2 or NDR1/2 kinases.
- Antibody validation: Serve as a positive control antigen for validating anti-FRYL antibodies in Western blot, ELISA, or immunofluorescence experiments.
- Structural studies: Suitable for use in structural biology workflows including cryo-EM or X-ray crystallography studies of FRYL domains or complexes (subject to protein construct and purity requirements).
- Epigenetic and chromatin research: Investigate FRYL's putative role in chromatin-associated complexes and transcriptional regulation using recombinant protein in in vitro reconstitution assays.
Researchers are encouraged to optimize assay conditions (buffer, pH, salt concentration, cofactors) empirically for their specific experimental system.
Troubleshooting Common Issues
When working with Recombinant Human FRYL Protein, researchers may encounter the following challenges. The guidance below provides starting points for troubleshooting:
| Issue | Possible Cause | Suggested Action |
|---|---|---|
| Low or no protein activity | Degradation from improper storage or excess freeze-thaw cycles | Use freshly thawed aliquots; confirm storage conditions; check CoA for activity data |
| Protein precipitation or aggregation | Incompatible buffer conditions or high protein concentration | Dilute in recommended buffer; add 0.1% BSA; centrifuge at 10,000 × g before use |
| Unexpected band size on SDS-PAGE | Glycosylation, post-translational modifications, or construct design | Compare with CoA reference gel image; consult Aladdin Scientific technical support |
| Weak signal in binding assays | Suboptimal assay buffer or protein concentration | Optimize salt concentration, pH, and protein input; include positive controls |
Frequently asked questions
What is the FRYL protein and why is it important for research?
FRYL (FRY-like transcription coactivator) is a large scaffold protein involved in NDR kinase signaling, the Hippo pathway, cell polarity, and transcriptional coactivation. It is important for research because of its roles in controlling tissue growth, organ size, and potentially epigenetic regulation, making it relevant to cancer biology, developmental biology, and chromatin research.
How should I store Recombinant Human FRYL Protein to maintain its activity?
Store Recombinant Human FRYL Protein at –80 °C for long-term preservation. Prepare single-use aliquots upon first thaw to avoid repeated freeze-thaw cycles, which can cause aggregation and loss of biological activity. Always refer to the specific storage conditions stated on the product datasheet and Certificate of Analysis.
What applications is Recombinant Human FRYL Protein suitable for?
Recombinant Human FRYL Protein (rp373177) is suitable for protein–protein interaction studies (Co-IP, pull-down assays, SPR), kinase activity assays, antibody validation as a positive control antigen, structural biology experiments, and in vitro reconstitution assays for chromatin and epigenetic research. Researchers should optimize assay conditions for their specific experimental system.
How do I reconstitute lyophilized Recombinant Human FRYL Protein?
If the product is supplied in lyophilized form, reconstitute it by adding the volume of buffer specified on the datasheet. Allow the protein to dissolve gently at room temperature or on ice without vigorous vortexing to prevent denaturation. After reconstitution, aliquot immediately and store at –80 °C.
Can Recombinant Human FRYL Protein be used for structural biology studies?
Recombinant Human FRYL Protein may be applicable to structural biology workflows such as cryo-EM or X-ray crystallography, depending on the specific protein construct, purity level, and homogeneity of the preparation. Researchers should verify that the purity and formulation specifications meet the requirements of their structural biology platform before use.
References
- Fang X, et al. Furry proteins in cell polarity, morphogenesis, and the regulation of NDR kinases. Cell Cycle. General textbook reference on FRY/FRYL family proteins and NDR kinase regulation.
- Harvey KF, Hariharan IK. The hippo pathway. Cold Spring Harbor Perspectives in Biology. General reference for Hippo pathway context including NDR kinase and scaffold protein interactions.
- Aladdin Scientific Product Datasheet: Recombinant Human FRYL Protein, SKU rp373177. Aladdin Scientific (absea#PP-7097).
Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.