Recombinant Human MED10 Protein: Research Guide & Applications

Recombinant Human MED10 Protein is a key subunit of the Mediator transcriptional coactivator complex, produced recombinantly for use in biochemical, structural, and gene-expression research applications.

Product Name

Recombinant Human MED10 Protein

SKU

rp367595

Protein Family

Mediator complex subunit (MED10)

Species Origin

Human (Homo sapiens)

Primary Application

Transcription regulation research, protein-protein interaction studies, biochemical assays

Format

Recombinant protein (lyophilized or liquid; confirm with datasheet)

Biological Background of MED10

MED10 (Mediator of RNA polymerase II transcription subunit 10) is an integral component of the Mediator complex, a large multiprotein assembly that functions as a transcriptional coactivator bridging gene-specific transcription factors and the general RNA polymerase II (Pol II) transcription machinery. The Mediator complex is essential for the regulated transcription of nearly all Pol II-dependent genes in eukaryotes.

MED10 is part of the core module of the Mediator complex and contributes to the structural integrity and functional activity of the complex. Through its participation in Mediator, MED10 is involved in the signal-dependent activation and repression of transcription, playing roles in cell differentiation, development, and homeostasis. Dysregulation of Mediator subunits, including MED10, has been implicated in various disease states, making recombinant MED10 a valuable research tool for understanding transcriptional control mechanisms.

Reconstitution and Handling Protocol

Reconstitution: If supplied in lyophilized form, briefly centrifuge the vial before opening to consolidate the contents. Reconstitute the protein in sterile, ultrapure water or an appropriate buffer (such as PBS, pH 7.4, supplemented with a carrier protein such as 0.1% BSA if required) to a working concentration recommended in the product datasheet. Allow the solution to equilibrate at room temperature for a few minutes with gentle mixing; do not vortex vigorously.

Aliquoting: To minimize freeze-thaw cycles and preserve activity, prepare single-use aliquots immediately after reconstitution. Avoid repeated freeze-thaw cycles, as these can lead to protein aggregation and loss of activity.

Handling: Work on ice when possible. Use low-protein-binding tubes and pipette tips to minimize adsorption losses. Refer to the official Aladdin Scientific product datasheet for lot-specific reconstitution volumes, concentrations, and buffer compositions.

Note: Specific reconstitution volumes and buffer details should always be confirmed against the current product datasheet provided with each lot (referencing absea#PP-3022 or SKU rp367595).

Storage and Stability Guidelines

Short-term storage: For use within 1–2 weeks, the reconstituted protein may be stored at 4 °C. Ensure the vial is tightly sealed and stored upright to prevent contamination and evaporation.

Long-term storage: For extended storage, aliquot the reconstituted protein and store at −20 °C or −80 °C. Storage at −80 °C is generally preferred for maximum stability and to maintain biological activity over longer periods.

Avoid: Repeated freeze-thaw cycles, exposure to temperatures above the recommended range, and prolonged storage at 4 °C. Addition of a carrier protein (e.g., 0.1% BSA) may enhance stability during storage at low concentrations.

Note: Always consult the product-specific datasheet (rp367595) for validated storage conditions, as formulation details may vary by lot.

Research Applications

Recombinant Human MED10 Protein is a versatile tool for a range of biochemical and cell-biology research applications:

• Protein–protein interaction studies: MED10 can be used in pull-down assays, co-immunoprecipitation experiments, and surface plasmon resonance (SPR) studies to characterize interactions within the Mediator complex or with transcription factors.
• Structural biology: Recombinant MED10 is suitable as a component for in vitro reconstitution of Mediator sub-complexes to support cryo-EM, X-ray crystallography, or other structural analyses.
• Biochemical assays: Useful as a positive control or standard in Western blotting, ELISA, and activity assays related to transcriptional regulation.
• Antibody production and validation: High-purity recombinant MED10 can serve as an immunogen for antibody generation or as an antigen standard for antibody validation.
• Gene expression and transcription studies: Supports investigation of Mediator-dependent transcriptional programs relevant to development, cancer biology, and metabolic disease.

Quality Attributes and Purity Considerations

Recombinant Human MED10 Protein (rp367595) is produced using recombinant expression systems to ensure consistency and reproducibility across experiments. Key quality attributes to verify on the product datasheet include:

• Purity: Purity level as assessed by SDS-PAGE or HPLC (typically ≥85–95% for research-grade recombinant proteins; confirm lot-specific value on the datasheet).
• Endotoxin level: Endotoxin content should be low (typically <1.0 EU/µg) for applications requiring cell-based assays; confirm on the Certificate of Analysis (CoA).
• Biological activity: Where applicable, functional activity may be confirmed by binding assays or transcription-related in vitro assays.
• Molecular weight: Confirm the expected molecular weight (accounting for any affinity tags) on the datasheet for rp367595.

Always request the Certificate of Analysis (CoA) for the specific lot received to verify these quality parameters prior to use in critical experiments.

Frequently asked questions

What is the biological function of MED10 in human cells?

MED10 is a subunit of the Mediator complex, a large multiprotein assembly that acts as a transcriptional coactivator. It bridges specific transcription factors and the RNA polymerase II transcription machinery, enabling regulated expression of Pol II-dependent genes involved in development, differentiation, and cellular homeostasis.

How should Recombinant Human MED10 Protein be reconstituted?

If supplied in lyophilized form, centrifuge the vial briefly before opening, then reconstitute in sterile ultrapure water or an appropriate buffer such as PBS (pH 7.4), optionally supplemented with 0.1% BSA as a carrier. Mix gently and avoid vigorous vortexing. Always refer to the lot-specific datasheet for exact reconstitution instructions for rp367595.

What are the recommended storage conditions for Recombinant Human MED10 Protein?

For short-term use (within 1–2 weeks), store reconstituted protein at 4 °C in a sealed, upright vial. For long-term storage, aliquot and store at −20 °C or preferably −80 °C to maintain stability and activity. Avoid repeated freeze-thaw cycles. Consult the product datasheet for lot-specific storage recommendations.

What research applications is Recombinant Human MED10 Protein suitable for?

Recombinant Human MED10 Protein (rp367595) is suitable for protein–protein interaction studies (pull-down, co-IP, SPR), structural biology (in vitro reconstitution of Mediator sub-complexes), Western blotting, ELISA, antibody production and validation, and investigation of transcriptional regulation mechanisms.

How can I verify the purity and quality of the recombinant protein I receive?

Request the Certificate of Analysis (CoA) for your specific lot of rp367595 from Aladdin Scientific. The CoA should provide purity data (SDS-PAGE or HPLC), endotoxin levels, and any functional activity results. Compare these values against the stated specifications on the product datasheet before use in critical experiments.

References

1. Tsai KL, Sato S, Tomomori-Sato C, Conaway RC, Conaway JW, Bhatt AN. A conserved Mediator-CDK8 kinase module association regulates Mediator-RNA polymerase II interaction. Nature Structural & Molecular Biology. 2013.
2. Allen BL, Taatjes DJ. The Mediator complex: a central integrator of transcription. Nature Reviews Molecular Cell Biology. 2015;16(3):155–166.
3. Aladdin Scientific. Recombinant Human MED10 Protein (rp367595) Product Datasheet. Aladdin Scientific; [current edition].

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