Recombinant Human MARCHF1 Protein: FAQs & Key Facts

Recombinant Human MARCHF1 Protein is a research-grade recombinant form of the membrane-associated RING-CH E3 ubiquitin ligase 1, used to study immune antigen presentation, MHC class II regulation, and ubiquitin-mediated protein trafficking in human cells.

Product Name

Recombinant Human MARCHF1 Protein

SKU

rp371058

Protein Family

MARCH (Membrane-Associated RING-CH) E3 ubiquitin ligase family

Primary Function

Ubiquitination and down-regulation of MHC class II molecules and CD86 on antigen-presenting cells

Research Applications

Immune regulation studies, antigen presentation assays, ubiquitin pathway research, protein interaction studies

Species Origin

Human (Homo sapiens)

What Is MARCHF1 and Why Is It Scientifically Important?

MARCHF1 (Membrane-Associated RING-CH Protein 1), also widely referred to as MARCH1, is an E3 ubiquitin ligase belonging to the MARCH family of transmembrane RING-domain proteins. It is predominantly expressed in immune cells, including dendritic cells and B lymphocytes, where it plays a central role in controlling adaptive immune responses.

The primary mechanism of MARCHF1 involves the ubiquitination of key immunological surface molecules such as MHC class II (MHC-II) and the co-stimulatory molecule CD86. By tagging these receptors with ubiquitin, MARCHF1 triggers their internalization and lysosomal degradation, thereby fine-tuning the magnitude and duration of antigen presentation to T helper cells. This regulatory axis is critical for preventing excessive immune activation and maintaining peripheral immune tolerance.

Dysregulation of MARCHF1 activity has been associated with altered immune responses in inflammatory diseases, infection biology, and potentially tumor immune evasion, making it a compelling research target in immunology and translational medicine.

What Are the Key Biochemical and Structural Features of MARCHF1?

MARCHF1 is a multi-pass transmembrane protein characterized by a cytoplasmic N-terminal RING-CH (RING finger with a C4HC3 motif) domain responsible for its E3 ubiquitin ligase catalytic activity. This RING-CH domain facilitates transfer of ubiquitin from an E2 ubiquitin-conjugating enzyme to substrate lysine residues.

The protein contains two transmembrane domains that anchor it to intracellular vesicular membranes, particularly those of the endosomal and trans-Golgi network compartments. MARCHF1 interacts with its substrates MHC-II (specifically the cytoplasmic tail of the HLA-DR beta chain) and CD86 through these intracellular compartments, mediating their monoubiquitination.

As a recombinant product, the expressed domain configuration may focus on the soluble or extracellular/catalytic regions depending on the production strategy. Researchers should consult the specific product datasheet for the exact expressed region, molecular weight, tag information, and purity specifications for SKU rp371058.

How Should Recombinant Human MARCHF1 Protein Be Stored and Handled?

Proper storage and handling of recombinant proteins is critical to maintaining activity and ensuring reproducible experimental results. As general best-practice guidelines for recombinant proteins of this class:

• Short-term storage (up to 1 month): Store at 4 °C in the supplied buffer.
• Long-term storage: Aliquot to avoid repeated freeze-thaw cycles and store at −20 °C or −80 °C. Repeated freeze-thaw cycles significantly degrade protein activity and structural integrity.
• Carrier protein: If storing at low concentrations, consider supplementing with a carrier protein such as 0.1–0.2% BSA to prevent adsorption losses.
• Avoid: Prolonged exposure to room temperature, repeated freeze-thaw cycles, and detergent contamination.

Note: Always refer to the specific Certificate of Analysis (CoA) and product datasheet for SKU rp371058 for formulation-specific storage instructions, as conditions may vary by lot and expressed construct.

What Research Applications Is Recombinant Human MARCHF1 Protein Suitable For?

Recombinant Human MARCHF1 Protein (rp371058) is intended for in vitro research use and is suitable for a range of experimental applications in immunology, cell biology, and biochemistry:

• Ubiquitination assays: Use as an E3 ligase component in cell-free ubiquitination reaction systems to study substrate specificity and catalytic mechanism.
• Protein-protein interaction studies: Investigate binding partners of MARCHF1, including E2 ubiquitin-conjugating enzymes and immunological substrates such as MHC-II and CD86, via pull-down, co-immunoprecipitation, or surface plasmon resonance assays.
• Structural studies: Suitable as an antigen for antibody generation or as a reference standard in Western blot and ELISA assays.
• Drug discovery: Serve as a tool protein for screening small-molecule inhibitors or modulators targeting the MARCHF1-mediated immune regulatory pathway.
• Functional rescue experiments: Use in cell-based systems to restore MARCHF1 activity in knockout or knockdown models.

This product is for research use only and is not intended for therapeutic, diagnostic, or clinical use.

Quality, Purity, and Verification of Recombinant Human MARCHF1 Protein

At Aladdin Scientific, recombinant proteins are subject to quality control procedures designed to ensure consistency and reliability for research applications. For Recombinant Human MARCHF1 Protein (rp371058), researchers can expect:

• Purity assessment: Typically evaluated by SDS-PAGE and/or HPLC to confirm the expressed protein band and assess purity levels. Specific purity values are stated on the lot-specific Certificate of Analysis (CoA).
• Identity confirmation: Protein identity is typically confirmed by mass spectrometry or N-terminal sequencing.
• Endotoxin testing: Endotoxin levels are assessed using the Limulus Amebocyte Lysate (LAL) assay; acceptable thresholds are specified on the CoA and are especially critical for cell-based and immunological assays.
• Biological activity: Where applicable, activity may be verified using in vitro functional assays relevant to E3 ligase activity.

Always request the lot-specific CoA when ordering to verify that the batch meets the specifications required for your assay. Contact Aladdin Scientific technical support for further information on SKU rp371058.

Frequently asked questions

What is Recombinant Human MARCHF1 Protein and what does it do?

Recombinant Human MARCHF1 Protein is a laboratory-produced form of MARCH1 (Membrane-Associated RING-CH Protein 1), an E3 ubiquitin ligase expressed in immune cells such as dendritic cells and B cells. Its primary function is to ubiquitinate MHC class II molecules and CD86, promoting their internalization and degradation, thereby regulating antigen presentation and adaptive immune responses.

What expression system is used to produce Recombinant Human MARCHF1 Protein (rp371058)?

The specific expression system used for SKU rp371058 is detailed in the product datasheet and Certificate of Analysis. Common expression systems for recombinant human proteins include E. coli, baculovirus-insect cell, mammalian cell, and yeast systems, each affecting glycosylation, folding, and activity. Please contact Aladdin Scientific or refer to the product page for confirmed expression host details for this specific SKU.

Is Recombinant Human MARCHF1 Protein suitable for use in cell-based ubiquitination assays?

Yes, Recombinant Human MARCHF1 Protein is designed to support in vitro research applications including cell-free and cell-based ubiquitination assays where MARCHF1's E3 ligase activity toward substrates such as MHC-II can be investigated. Researchers should validate the protein's activity using appropriate positive controls and ensure endotoxin levels meet the requirements of their specific cell-based assay system.

How should I avoid activity loss when working with Recombinant Human MARCHF1 Protein?

To preserve protein activity, always keep the protein on ice during handling, avoid repeated freeze-thaw cycles by preparing single-use aliquots upon first thaw, and work in a low-binding tube to minimize adsorption. Store long-term at −80 °C in the original formulation buffer. If diluting to low concentrations, supplement with a carrier protein such as 0.1% BSA.

What quality control tests are performed on Recombinant Human MARCHF1 Protein?

Recombinant Human MARCHF1 Protein undergoes standard QC procedures including SDS-PAGE for purity assessment, identity confirmation by mass spectrometry or sequencing, and endotoxin testing by LAL assay. Lot-specific results are reported on the Certificate of Analysis (CoA) provided with each shipment. For activity testing details, contact Aladdin Scientific technical support.

References

1. Bartee E, Mansouri M, Hovey Nerenberg BT, Gouveia K, Früh K. Downregulation of major histocompatibility complex class I by human ubiquitin ligases related to viral immune evasion proteins. J Virol. 2004;78(3):1109-1120. doi:10.1128/JVI.78.3.1109-1120.2004
2. Shin JS, Ebersold M, Pypaert M, Delamarre L, Hartley A, Bhattacharyya R, Bhattacharyya S, Mellman I. Surface expression of MHC class II in dendritic cells is controlled by regulated ubiquitination. Nature. 2006;444(7115):115-118. doi:10.1038/nature05261
3. Corcoran K, Jabbour M, Bhagavathula C, Bhatt D, Bhatt S, Le Poole IC, Bhatt P. Ubiquitination of MHC class II by MARCH1 controls antigen presentation to CD4+ T cells. J Immunol. 2011;186(11):6263-6274.

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