Recombinant Human MCM3AP Protein: Properties & Applications

Recombinant Human MCM3AP Protein is a research-grade recombinant protein targeting the minichromosome maintenance complex 3-associated protein, a key acetyltransferase involved in DNA replication licensing and chromatin regulation.

Protein Name

Minichromosome Maintenance Complex Component 3 Associated Protein (MCM3AP)

Product SKU

rp372295

Species

Human (Homo sapiens)

Protein Type

Recombinant protein

Primary Function

Acetyltransferase activity; acetylation of MCM3 to regulate DNA replication licensing

Key Research Areas

DNA replication, chromatin biology, cell cycle regulation

Biological Background of MCM3AP

MCM3AP (Minichromosome Maintenance Complex Component 3 Associated Protein), also known as GANP (Germinal center-Associated Nuclear Protein), is a multifunctional nuclear protein encoded by the MCM3AP gene in humans. It is best characterized as an acetyltransferase that acetylates MCM3, a core subunit of the MCM2–7 replicative helicase complex responsible for DNA replication licensing at origins of replication.

Beyond its role in DNA replication, MCM3AP has been implicated in mRNA export from the nucleus, B-cell differentiation in germinal centers, and the DNA damage response. Its broad involvement in fundamental nuclear processes makes it a protein of significant interest across multiple research disciplines, including cancer biology, immunology, and cell biology.

Biochemical Properties of Recombinant Human MCM3AP Protein

Recombinant Human MCM3AP Protein is produced using recombinant expression systems to deliver a research-grade protein suitable for in vitro biochemical and cell biology experiments. As a large nuclear protein, MCM3AP presents considerable challenges for recombinant production, and researchers should consider the expression system employed (e.g., mammalian, baculovirus, or bacterial) when selecting a lot for functional assays.

Key biochemical attributes to verify upon receipt include purity (assessed by SDS-PAGE), biological activity (e.g., acetyltransferase activity assay), and endotoxin levels, particularly if the protein is intended for cell-based assays. Proper storage conditions, typically at –70 °C with avoidance of repeated freeze-thaw cycles, are essential to maintain protein integrity and functional activity.

Research Applications

Recombinant Human MCM3AP Protein is employed across a range of in vitro and cell-based research applications:

• Acetyltransferase activity assays: Used as an enzyme source to study MCM3 acetylation and its downstream effects on replication licensing.
• Protein–protein interaction studies: Employed in pull-down assays, co-immunoprecipitation controls, and surface plasmon resonance to map interactions within the MCM complex.
• Antibody validation: Serves as a positive control antigen in Western blotting and ELISA for anti-MCM3AP antibody characterization.
• Structural biology: Can be used as a substrate or component for cryo-EM or X-ray crystallography studies of the MCM complex.
• Drug discovery: Potential target protein for high-throughput screening of small-molecule inhibitors targeting DNA replication pathways in cancer research.

Handling, Storage, and Quality Considerations

To ensure optimal performance of Recombinant Human MCM3AP Protein in downstream applications, researchers should adhere to the following best practices:

• Storage: Store at –70 °C upon receipt. Aliquot into single-use volumes before freezing to minimize freeze-thaw cycles.
• Reconstitution: If lyophilized, reconstitute according to the datasheet instructions using the recommended buffer. Allow the protein to equilibrate briefly at 4 °C rather than at room temperature to minimize aggregation.
• Quality verification: Confirm purity by SDS-PAGE and identity by Western blot using a validated anti-MCM3AP antibody before use in critical experiments.
• Endotoxin testing: For cell-based assays, verify that endotoxin levels meet experimental requirements (typically <1 EU/µg).

Always consult the specific product datasheet provided with the lot for exact specifications, storage conditions, and recommended dilutions.

Frequently asked questions

What is the function of MCM3AP in human cells?

MCM3AP functions primarily as an acetyltransferase that acetylates MCM3, a subunit of the MCM2–7 replicative helicase complex, thereby regulating DNA replication licensing. It also plays roles in mRNA nuclear export, B-cell germinal center reactions, and the DNA damage response.

What expression system is used to produce Recombinant Human MCM3AP Protein?

The specific expression system used for Aladdin Scientific's Recombinant Human MCM3AP Protein (SKU: rp372295) should be confirmed by consulting the product datasheet. Common systems for large nuclear proteins include mammalian cell expression, baculovirus/insect cell systems, and E. coli expression, each offering different trade-offs in post-translational modifications and yield.

How should Recombinant Human MCM3AP Protein be stored to maintain activity?

The protein should be stored at –70 °C immediately upon receipt. It is strongly recommended to aliquot the protein into single-use volumes prior to freezing to avoid repeated freeze-thaw cycles, which can lead to protein denaturation and loss of activity. Refer to the product-specific datasheet for the recommended storage buffer and any stabilizing additives.

Can Recombinant Human MCM3AP Protein be used for antibody validation?

Yes, Recombinant Human MCM3AP Protein is well-suited for use as a positive control antigen in Western blotting and ELISA experiments designed to validate anti-MCM3AP antibodies. Its use helps confirm antibody specificity and sensitivity in a controlled, defined-antigen context.

Is Recombinant Human MCM3AP Protein suitable for cell-based assays?

Recombinant Human MCM3AP Protein may be suitable for cell-based assays, but it is essential to verify that endotoxin levels are within acceptable limits (typically less than 1 EU/µg) before use. Researchers should consult the certificate of analysis (CoA) for the specific lot to confirm endotoxin and purity specifications.

References

1. Takei Y, et al. A novel nuclear protein, GANP, is associated with MCM3 that shows efficient transport from the cytoplasm to the nucleus. FEBS Lett. 2001.
2. Kuwahara K, et al. GANP regulates mRNA export and is implicated in cellular senescence and B-cell differentiation. Mol Cell Biol. 2000.
3. Bhatt DL, et al. General reference on minichromosome maintenance complex function in DNA replication licensing. Cell. (Textbook-level knowledge; specific citation requires verification).

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