Recombinant Human MED20 Protein: Properties & Research Guide

Recombinant Human MED20 Protein is a key subunit of the Mediator transcriptional coactivator complex, produced recombinantly to support studies of RNA polymerase II-dependent gene regulation and protein–protein interaction networks in eukaryotic transcription.

Protein Name

Recombinant Human MED20 Protein

Gene Name

MED20 (also known as TRAP17, DRIP34)

Protein Family

Mediator complex, head module subunit

Biological Function

Structural and functional component of the Mediator complex; facilitates communication between gene-specific transcription factors and RNA polymerase II

Expression System

Recombinant expression (host system details to be confirmed from datasheet)

Research Applications

Transcription studies, protein interaction assays, structural biology, biochemical reconstitution

SKU

rp368434

Biological Background of MED20

MED20 is a subunit of the Mediator complex, an evolutionarily conserved multiprotein assembly that serves as a central regulator of RNA polymerase II (Pol II)-dependent transcription in eukaryotes. The Mediator complex bridges sequence-specific transcriptional activators and repressors bound at enhancers or promoters with the general transcription machinery assembled at the core promoter.

MED20 localizes to the head module of the Mediator complex, which makes direct contact with the C-terminal domain (CTD) of the largest subunit of RNA polymerase II. This positioning makes MED20 integral to the initiation of transcription and to the regulation of diverse gene expression programs in human cells. Dysregulation of Mediator subunits, including MED20, has been associated with developmental abnormalities and oncogenic transformation, making these proteins subjects of active biomedical research.

Biochemical Properties and Product Specifications

Recombinant Human MED20 Protein (SKU: rp368434) is produced using recombinant expression technology to deliver a well-characterized protein suitable for laboratory research. The following points summarize key biochemical considerations for this product:

• Origin: Human (Homo sapiens), gene MED20.
• Protein form: Recombinant, typically supplied as a lyophilized powder or liquid formulation; confirm format with the product datasheet.
• Purity: Recombinant proteins in this category are generally produced to >85% purity as assessed by SDS-PAGE; verify exact purity specification on the Certificate of Analysis (CoA).
• Formulation: Buffer composition and stabilizers should be confirmed from the product-specific datasheet or CoA prior to use.
• Storage: Typically stored at -20°C or -80°C; avoid repeated freeze-thaw cycles to maintain activity and integrity.

Note: Specific molecular weight, tag information, expression host, and endotoxin levels should be verified against the current product datasheet for SKU rp368434.

Recommended Handling and Reconstitution Protocol

Proper handling of Recombinant Human MED20 Protein is essential for preserving its structural integrity and functional activity. The following general protocol is recommended; always cross-reference with the product-specific datasheet.

1. Thawing: Thaw the protein on ice or at 4°C. Do not thaw at room temperature, as this may cause aggregation or degradation.
2. Reconstitution (lyophilized product): Reconstitute with the volume of sterile water or appropriate buffer as specified on the datasheet. Gently mix by pipetting; avoid vortexing.
3. Centrifugation: Briefly centrifuge the vial (1,000–2,000 × g, 30 seconds) before opening to consolidate contents.
4. Aliquoting: Prepare single-use aliquots immediately after reconstitution to avoid repeated freeze-thaw cycles.
5. Storage after reconstitution: Use immediately or store at -80°C for long-term. At 4°C, stability is typically limited to a few days.
6. Carrier protein: If the protein is used at very low concentrations, consider adding a carrier protein (e.g., 0.1% BSA) to prevent adsorption to tube surfaces.

Always consult the Certificate of Analysis and Safety Data Sheet (SDS) before use. Handle recombinant proteins with appropriate personal protective equipment (PPE).

Research Applications

Recombinant Human MED20 Protein supports a range of experimental applications in molecular biology and biochemistry research:

• Protein–protein interaction studies: Used in pull-down assays, co-immunoprecipitation (co-IP), or surface plasmon resonance (SPR) to characterize MED20 interactions with other Mediator subunits and transcription factors.
• Structural biology: Suitable as a component for cryo-electron microscopy (cryo-EM) or X-ray crystallography studies aimed at resolving the architecture of the Mediator head module.
• Biochemical reconstitution: Enables in vitro reconstitution of partial or full Mediator complexes for functional transcription assays.
• Antibody development and validation: Can serve as an antigen standard for raising or validating antibodies targeting MED20.
• Drug discovery: Useful in screening assays designed to identify small molecules that modulate Mediator–transcription factor interactions relevant to oncology or developmental biology.

These applications make Recombinant Human MED20 Protein a versatile reagent for laboratories focused on transcriptional regulation, epigenetics, and therapeutic target identification.

Quality, Storage, and Regulatory Considerations

To ensure experimental reproducibility and reagent integrity, observe the following quality and regulatory guidelines when working with Recombinant Human MED20 Protein:

• Lot-to-lot consistency: Request a Certificate of Analysis (CoA) for each lot to confirm purity, concentration, and functional test results before use in critical experiments.
• Endotoxin levels: For cell-based assays, confirm that endotoxin levels are within acceptable limits as stated on the CoA; elevated endotoxin can confound transcriptional assays.
• Research use only: Recombinant Human MED20 Protein (rp368434) is intended for research use only and is not approved for diagnostic or therapeutic applications in humans.
• Shipping conditions: Recombinant proteins are typically shipped on dry ice; inspect the package upon receipt and document any anomalies immediately.
• Regulatory compliance: Dispose of protein-containing materials in accordance with institutional biosafety guidelines and local regulations.

Frequently asked questions

What is MED20 and why is it important in transcription research?

MED20 is a subunit of the Mediator complex, a conserved multiprotein assembly essential for RNA polymerase II-dependent transcription in eukaryotes. It resides in the head module of Mediator and helps relay signals from transcriptional activators to the core transcription machinery, making it central to understanding gene regulation, development, and disease mechanisms including cancer.

How should Recombinant Human MED20 Protein be stored to maintain stability?

Recombinant Human MED20 Protein should be stored at -20°C or preferably -80°C for long-term stability. Repeated freeze-thaw cycles should be avoided by preparing single-use aliquots after reconstitution or upon first thaw. Always refer to the product-specific datasheet and Certificate of Analysis for the confirmed storage conditions for lot rp368434.

What experimental applications is Recombinant Human MED20 Protein validated for?

Recombinant Human MED20 Protein is suitable for use in protein–protein interaction studies (pull-down, co-IP, SPR), structural biology (cryo-EM, crystallography), biochemical reconstitution of Mediator complexes, antibody development and validation, and drug discovery screening assays. Specific validated applications should be confirmed on the product datasheet for SKU rp368434.

Is Recombinant Human MED20 Protein suitable for use in cell-based assays?

Recombinant Human MED20 Protein may be used in cell-based experimental contexts, but it is critical to verify the endotoxin level reported on the Certificate of Analysis before use, as elevated endotoxin contamination can significantly interfere with transcriptional and cell signaling assays. This product is intended for research use only and is not approved for clinical applications.

How do I reconstitute lyophilized Recombinant Human MED20 Protein?

Briefly centrifuge the vial to consolidate contents, then add the volume of sterile water or appropriate buffer specified on the datasheet. Gently mix by pipetting without vortexing. Prepare single-use aliquots immediately to prevent repeated freeze-thaw cycles. The exact reconstitution buffer and volume are specified in the product datasheet for SKU rp368434.

References

1. Kelleher RJ 3rd, Flanagan PM, Kornberg RD. A novel mediator between activator proteins and the RNA polymerase II transcription apparatus. Cell. 1990;61(7):1209-1215.
2. Mittler G, Stühler T, Santolin L, et al. A novel docking site on Mediator is critical for activation by VP16 in mammalian cells. EMBO J. 2003;22(24):6494-6504.
3. Tsai KL, Sato S, Tomomori-Sato C, et al. A conserved Mediator-CDK8 kinase module association regulates Mediator-RNA polymerase II interaction. Nat Struct Mol Biol. 2013;20(5):611-619.

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