Recombinant Human CNOT4 Protein: FAQs & Key Facts

Recombinant Human CNOT4 Protein is a key research reagent enabling scientists to investigate the CCR4-NOT deadenylase complex, E3 ubiquitin ligase activity, and mRNA decay pathways in human cellular systems.

Product Name

Recombinant Human CNOT4 Protein

SKU

rp372206

Protein Function

Component of the CCR4-NOT complex; possesses E3 ubiquitin ligase activity via its RING finger domain

Biological Role

Involved in mRNA deadenylation, transcriptional regulation, and protein ubiquitination

Species

Human (Homo sapiens)

Format

Recombinant protein produced by heterologous expression

Supplier

Aladdin Scientific

What Is CNOT4 and Why Is It Important?

CNOT4 (CCR4-NOT Transcription Complex Subunit 4) is a multifunctional protein and a subunit of the evolutionarily conserved CCR4-NOT complex, one of the major cytoplasmic deadenylase complexes in eukaryotes. The CCR4-NOT complex plays a central role in mRNA decay, translational repression, and the regulation of gene expression at the post-transcriptional level.

CNOT4 itself is notable for harboring a RING (Really Interesting New Gene) finger domain in its N-terminal region, which confers E3 ubiquitin ligase activity. This means CNOT4 can facilitate the transfer of ubiquitin to target substrate proteins, marking them for proteasomal degradation or altering their function. Through this dual role—as a scaffold within the CCR4-NOT complex and as an independent E3 ligase—CNOT4 participates in the regulation of transcription, mRNA stability, protein turnover, and cellular stress responses.

Dysregulation of CNOT4 and the CCR4-NOT complex has been associated with developmental defects, cancer, and other human diseases, making CNOT4 a scientifically valuable target for biochemical and cell biology research.

What Are the Key Applications of Recombinant Human CNOT4 Protein?

Recombinant Human CNOT4 Protein (SKU: rp372206) is suitable for a range of in vitro and cell-free research applications, including:

• Biochemical assays: Use as a substrate or active enzyme in ubiquitination assays to characterize CNOT4 E3 ligase activity and identify potential substrates or inhibitors.
• Protein–protein interaction studies: Investigate how CNOT4 interacts with other CCR4-NOT subunits or partner proteins using pull-down, co-immunoprecipitation, or surface plasmon resonance (SPR) approaches.
• Structural studies: Provide purified recombinant protein for crystallography or cryo-electron microscopy studies of the CCR4-NOT complex architecture.
• Antibody production and validation: Use as an antigen for generating or validating antibodies specific to human CNOT4.
• Drug discovery: Screen small-molecule compound libraries for inhibitors or modulators of CNOT4 E3 ligase activity as potential therapeutic leads in oncology or other disease areas.

Researchers should confirm suitability for each specific application and optimize conditions empirically, as performance may vary by experimental context.

Storage, Handling, and Stability Guidelines

Proper storage and handling of Recombinant Human CNOT4 Protein are essential to maintain protein integrity and biological activity. General best-practice guidelines for recombinant proteins apply:

• Storage temperature: Store at −80 °C for long-term preservation. Avoid repeated freeze–thaw cycles, which can lead to protein aggregation and loss of activity.
• Short-term storage: If the protein will be used within a few days, storage at 4 °C may be acceptable; consult the product datasheet for confirmed conditions.
• Aliquoting: Upon receipt, aliquot the protein into single-use volumes to minimize freeze–thaw cycles.
• Carrier protein: Many recombinant proteins are stabilized with BSA or glycerol. Check the product-specific Certificate of Analysis (CoA) for formulation details.
• Avoid contamination: Use sterile, low-binding microtubes and pipette tips. Work on ice during handling sessions.

Note: Specific storage conditions, buffer composition, concentration, and stability data for SKU rp372206 should be verified against the current product datasheet and CoA supplied with each lot.

Quality, Purity, and Regulatory Considerations

Aladdin Scientific's Recombinant Human CNOT4 Protein is manufactured as a research-grade reagent intended for in vitro research use only. Key quality considerations include:

• Purity: Recombinant proteins in this product category are typically analyzed by SDS-PAGE to confirm purity. Specific purity data for rp372206 should be confirmed on the product's CoA.
• Identity verification: Product identity may be confirmed by Western blot or mass spectrometry. Refer to the CoA for lot-specific results.
• Endotoxin levels: For applications sensitive to endotoxin (e.g., cell-based assays), verify endotoxin content on the CoA. Low-endotoxin preparations may be required.
• Not for clinical or diagnostic use: This product is strictly for laboratory research purposes and is not approved for use in humans, animals, or as a diagnostic reagent.
• Lot-to-lot consistency: Contact Aladdin Scientific technical support for information on lot-specific quality documentation and consistency data.

Frequently asked questions

What is the biological function of CNOT4 in human cells?

CNOT4 is a subunit of the CCR4-NOT complex, the major mRNA deadenylase complex in eukaryotes, and plays roles in mRNA decay and transcriptional regulation. Its N-terminal RING finger domain endows it with E3 ubiquitin ligase activity, enabling it to ubiquitinate target proteins and regulate their stability or function. Through these activities, CNOT4 influences gene expression at both post-transcriptional and post-translational levels.

What expression system is used to produce Recombinant Human CNOT4 Protein (rp372206)?

The specific expression system (e.g., E. coli, baculovirus/insect cells, or mammalian cells) used for SKU rp372206 is not detailed in the currently available source material. Researchers requiring this information should consult the product datasheet or contact Aladdin Scientific technical support directly for lot-specific manufacturing details.

Is Recombinant Human CNOT4 Protein suitable for use in cell-based assays?

Recombinant Human CNOT4 Protein is primarily intended for in vitro biochemical applications. Use in cell-based assays may be possible but would require optimization of delivery method, concentration, and endotoxin levels. Researchers should verify endotoxin content on the Certificate of Analysis and determine empirically whether the protein is suitable for their specific cellular system.

How should I prevent loss of activity during routine use of CNOT4 recombinant protein?

To preserve protein activity, aliquot the stock immediately upon receipt to avoid repeated freeze–thaw cycles, store aliquots at −80 °C, and work on ice during experiments. Avoid exposure to detergents, extreme pH, or prolonged incubation at room temperature. Always refer to the product-specific datasheet for recommended handling conditions.

Can Recombinant Human CNOT4 Protein be used for structural biology studies?

Yes, purified recombinant proteins such as CNOT4 are commonly used in structural biology applications including X-ray crystallography and cryo-electron microscopy. However, suitability depends on purity, homogeneity, and the presence of tags that may need to be removed. Researchers should confirm lot-specific purity and formulation details with Aladdin Scientific before proceeding with structural studies.

References

1. Lau NC, et al. Human Ccr4-Not complexes contain variable deadenylase subunits. Biochemical Journal. 2009.
2. Hanzawa H, et al. The structure of the C4C4 RING finger of human CNOT4 reveals a novel zinc-binding domain. Nature Structural Biology. 2001.
3. Dölken L, et al. Transcriptional regulation and mRNA turnover via the CCR4-NOT complex. Molecular Cell. Review.

Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.