Recombinant Human MCFD2 Protein: Function & Research Use

Recombinant Human MCFD2 Protein is a critical research reagent that enables scientists to investigate the molecular mechanisms underlying combined coagulation factor V and VIII deficiency and ER-to-Golgi cargo transport.

Protein Name

Multiple Coagulation Factor Deficiency Protein 2 (MCFD2)

UniProt Accession

Q8NI22 (Human)

Protein Family

EF-hand calcium-binding protein family

Biological Function

Forms a cargo receptor complex with LMAN1 (ERGIC-53) to facilitate ER-to-Golgi transport of coagulation factors V and VIII

Associated Disease

Combined deficiency of coagulation factors V and VIII (F5F8D)

Expression System

Recombinant, expressed in E. coli or mammalian system (verify with datasheet)

SKU

rp369477

What Is MCFD2? Biological Background

MCFD2 (Multiple Coagulation Factor Deficiency Protein 2) is a soluble luminal protein encoded by the MCFD2 gene in humans. It is a member of the EF-hand calcium-binding protein family, characterized by two EF-hand motifs that mediate calcium-dependent interactions with its binding partner LMAN1 (also known as ERGIC-53), a type I transmembrane lectin resident in the endoplasmic reticulum–Golgi intermediate compartment (ERGIC).

The MCFD2–LMAN1 heteromeric complex functions as a cargo receptor that selectively captures the glycoprotein coagulation factors V (FV) and VIII (FVIII) in the ER lumen and packages them into COPII-coated vesicles for anterograde transport to the Golgi apparatus. This transport step is essential for the efficient secretion of FV and FVIII into the bloodstream.

Loss-of-function mutations in MCFD2 result in combined deficiency of coagulation factors V and VIII (F5F8D; OMIM #227300), a rare autosomal recessive bleeding disorder characterized by mildly to moderately reduced plasma levels of both FV and FVIII. Understanding the structure and function of MCFD2 is therefore of direct relevance to hemostasis research, rare bleeding disorder studies, and broader investigations into ER quality control and secretory pathway regulation.

Biochemical Properties of Recombinant Human MCFD2 Protein

Recombinant Human MCFD2 Protein (SKU: rp369477) is produced using recombinant DNA technology to deliver a highly pure, biologically relevant form of the human MCFD2 protein for use in laboratory research. Key biochemical characteristics include:

• Protein family: EF-hand calcium-binding protein; contains two canonical EF-hand calcium-binding motifs critical for LMAN1 interaction and cargo capture.
• Calcium dependence: The interaction between MCFD2 and LMAN1 is strictly calcium-dependent; EF-hand mutations that abolish calcium binding also abrogate complex formation and cargo transport.
• Molecular weight: The mature human MCFD2 protein has a predicted molecular weight of approximately 16 kDa, though apparent molecular weight on SDS-PAGE may differ depending on the tag and expression system used.
• Purity: Recombinant preparations are typically provided at high purity (consult the product datasheet for lot-specific purity data).
• Formulation: Supplied as a lyophilized powder or in solution; storage conditions and reconstitution buffer should be confirmed from the current product datasheet.

Note: Researchers should always refer to the current Aladdin Scientific product datasheet for lot-specific specifications including purity, activity, endotoxin level, and recommended storage conditions.

Research Applications

Recombinant Human MCFD2 Protein is a versatile tool for a range of biochemical and cell biology research applications:

• Protein–protein interaction studies: Investigating the calcium-dependent binding of MCFD2 to LMAN1 and to coagulation factor cargo (FV, FVIII) using techniques such as co-immunoprecipitation, surface plasmon resonance (SPR), isothermal titration calorimetry (ITC), and pull-down assays.
• Structural biology: As a substrate or binding partner for X-ray crystallography, cryo-EM, or NMR studies aimed at resolving the MCFD2–LMAN1 complex structure and EF-hand calcium coordination geometry.
• Functional assays: Reconstitution experiments to assess the role of wild-type or mutant MCFD2 in COPII vesicle packaging and secretion of FV/FVIII in cell-free or cell-based systems.
• Antibody validation and immunoassay development: Serving as a positive control antigen for Western blotting, ELISA, and immunofluorescence assays targeting MCFD2.
• Disease mechanism research: Characterizing the biochemical consequences of clinically identified MCFD2 missense and nonsense mutations associated with F5F8D.
• Drug discovery: Screening for small molecules that modulate the MCFD2–LMAN1 interaction as a potential therapeutic strategy for bleeding disorders or as a tool to understand secretory pathway biology.

Handling, Storage, and Quality Considerations

To maintain the integrity and activity of Recombinant Human MCFD2 Protein, researchers should observe the following best practices:

• Storage: Store according to the conditions specified on the product datasheet (typically –20 °C or –80 °C for long-term storage). Avoid repeated freeze–thaw cycles, which can compromise protein integrity.
• Reconstitution: If supplied as lyophilized powder, reconstitute in the buffer recommended on the datasheet, allow equilibration at room temperature for a few minutes, and gently mix without vortexing.
• Calcium supplementation: Since MCFD2 function is calcium-dependent, ensure that assay buffers contain appropriate concentrations of CaCl₂ when studying MCFD2–LMAN1 interactions or cargo-binding activities. EGTA or EDTA can be used as controls to confirm calcium dependence.
• Aliquoting: Prepare single-use aliquots upon first thaw to minimize freeze–thaw degradation.
• Endotoxin: For cell-based assays, verify endotoxin levels from the certificate of analysis (CoA) to avoid confounding inflammatory responses.
• Quality verification: Confirm protein identity and purity by SDS-PAGE and/or Western blot prior to critical experiments.

Always consult the current product datasheet and certificate of analysis supplied with each lot for authoritative specification data.

Ordering Information

Recombinant Human MCFD2 Protein is available from Aladdin Scientific. Please refer to the table below for ordering details:

SKU	Product Name	Supplier
rp369477	Recombinant Human MCFD2 Protein	Aladdin Scientific

For pricing, availability, bulk inquiries, or custom formulation requests, please contact Aladdin Scientific customer service or visit the product page on the Aladdin Scientific website. All products are intended for research use only and are not approved for diagnostic or therapeutic applications.

Frequently asked questions

What is the biological function of MCFD2?

MCFD2 forms a calcium-dependent cargo receptor complex with LMAN1 (ERGIC-53) in the endoplasmic reticulum lumen. This complex selectively binds coagulation factors V and VIII and packages them into COPII-coated transport vesicles for delivery to the Golgi apparatus, enabling their efficient secretion into the bloodstream.

What disease is associated with MCFD2 mutations?

Loss-of-function mutations in MCFD2 cause combined deficiency of coagulation factors V and VIII (F5F8D; OMIM #227300), a rare autosomal recessive bleeding disorder. Affected individuals have mildly to moderately reduced plasma levels of both FV and FVIII, leading to a bleeding tendency.

Why is calcium important for MCFD2 function, and how should I account for this in assays?

MCFD2 contains two EF-hand motifs that bind calcium ions, and calcium binding is essential for MCFD2 to interact with LMAN1 and capture coagulation factor cargo. When designing binding or functional assays, include CaCl₂ at physiologically relevant concentrations in your assay buffers, and use EGTA or EDTA as negative controls to confirm that observed interactions are calcium-dependent.

What applications is Recombinant Human MCFD2 Protein suitable for?

Recombinant Human MCFD2 Protein (rp369477) is suitable for protein–protein interaction studies, structural biology experiments, antibody validation (Western blot, ELISA), functional reconstitution assays, disease-associated mutation characterization, and drug discovery screening. Please consult the product datasheet for validated applications and recommended assay conditions.

How should Recombinant Human MCFD2 Protein be stored to maintain activity?

Store the protein according to the conditions specified on the product datasheet, typically at –20 °C or –80 °C for long-term storage. Avoid repeated freeze–thaw cycles by preparing single-use aliquots upon first thaw. Confirm specific storage and reconstitution instructions from the current datasheet provided with your lot.

References

1. Zhang B, Cunningham MA, Nichols WC, et al. Bleeding due to disruption of a cargo-specific ER-to-Golgi transport complex. Nature Genetics. 2003;34(2):220–228. doi:10.1038/ng1153
2. Nyfeler B, Kamber M, Vécsei A, et al. The cargo receptor MCFD2 is involved in the selective secretion of blood coagulation factors V and VIII. The EMBO Journal. 2005;24(8):1699–1709.
3. Kawasaki N, Ichikawa Y, Matsuo I, et al. The cargo receptor MCFD2 is a calcium-binding protein that interacts with LMAN1. Blood. 2008;111(3):1572–1579.
4. UniProt Consortium. UniProt entry Q8NI22 (MCFD2_HUMAN). UniProt Knowledgebase. Available at: https://www.uniprot.org/uniprot/Q8NI22

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