Recombinant Human GOLPH4 Protein: Properties & Applications

Recombinant Human GOLPH4 Protein is a research-grade reagent representing the human Golgi phosphoprotein 4, widely used to investigate Golgi apparatus organization, membrane trafficking pathways, and protein glycosylation mechanisms in cell biology research.

Protein Name

Recombinant Human GOLPH4 Protein

Also Known As

Golgi Phosphoprotein 4; GP73; GOLM1-related protein

Biological Origin

Human (Homo sapiens)

Product SKU

rp366300

Supplier Reference

absea#PP-1905

Primary Research Area

Golgi apparatus biology, membrane trafficking, protein secretion

Typical Applications

ELISA, Western blot, functional assays, antibody production

Biological Background of GOLPH4

GOLPH4, also referred to as Golgi Phosphoprotein 4, is a type II transmembrane protein that localizes predominantly to the Golgi apparatus—the central organelle responsible for protein sorting, modification, and trafficking within eukaryotic cells. The Golgi apparatus processes and directs proteins synthesized in the endoplasmic reticulum (ER) to their final destinations, including the cell surface, lysosomes, and the extracellular space.

GOLPH4 is structurally characterized by a short N-terminal cytoplasmic tail, a single transmembrane domain, and a large luminal domain. This topology is consistent with many Golgi-resident proteins that play structural or enzymatic roles within the Golgi stack. As a phosphoprotein, GOLPH4 may be subject to phosphorylation-dependent regulatory mechanisms, although the precise signaling pathways governing its activity remain an active area of investigation.

Research interest in GOLPH4 has grown due to its potential involvement in Golgi integrity, glycosylation reactions, and protein secretion. Understanding the molecular functions of GOLPH4 may provide insights into diseases associated with Golgi dysfunction, including certain cancers and congenital disorders of glycosylation.

Biochemical Properties and Production

Recombinant Human GOLPH4 Protein is produced using established recombinant expression systems designed to yield a soluble, functional form of the protein suitable for in vitro research applications. Recombinant proteins of this class are typically expressed in mammalian, insect (baculovirus), or prokaryotic (E. coli) host systems, each offering distinct advantages in terms of post-translational modifications, yield, and scalability.

For a Golgi-resident phosphoprotein such as GOLPH4, mammalian expression systems are often preferred when native glycosylation patterns and proper folding are critical for downstream assays. The recombinant protein is generally purified by affinity chromatography, often utilizing a His-tag or GST-tag appended to the recombinant construct, followed by additional polishing steps such as size-exclusion chromatography to ensure high purity and batch-to-batch consistency.

Quality control for recombinant proteins typically includes assessment of purity by SDS-PAGE, molecular weight confirmation, endotoxin testing, and functional validation. Researchers should consult the product datasheet for lot-specific information on purity, concentration, formulation buffer, and recommended storage conditions to ensure optimal experimental outcomes.

Research Applications

Recombinant Human GOLPH4 Protein is a versatile tool applicable across a range of experimental paradigms in cell and molecular biology. Key applications include:

• Enzyme-Linked Immunosorbent Assay (ELISA): Recombinant GOLPH4 can serve as a standard antigen for developing and validating ELISA-based immunoassays, enabling quantitative measurement of GOLPH4 in biological samples.
• Western Blot (WB): The purified protein is useful as a positive control in Western blotting experiments to confirm antibody specificity and sensitivity when probing for endogenous GOLPH4 in cell or tissue lysates.
• Antibody Production and Validation: High-purity recombinant GOLPH4 serves as an immunogen for generating monoclonal or polyclonal antibodies, as well as a reference standard for antibody characterization.
• Functional and Binding Assays: The protein may be employed in pull-down assays, surface plasmon resonance (SPR), or co-immunoprecipitation studies to identify interaction partners and elucidate the molecular mechanisms underlying GOLPH4's role in Golgi function and membrane trafficking.
• Structural Studies: Purified recombinant protein supports biophysical analyses including dynamic light scattering (DLS) or crystallography efforts aimed at resolving GOLPH4 protein structure.

These applications collectively support a broad research agenda spanning fundamental Golgi biology to translational studies linking Golgi dysfunction to human disease.

Handling, Storage, and Experimental Tips

Proper handling and storage of recombinant proteins are essential to preserve activity and ensure reproducible experimental results. The following general guidelines apply to Recombinant Human GOLPH4 Protein:

• Storage Temperature: Recombinant proteins should typically be stored at –20°C or –80°C for long-term stability. Avoid repeated freeze-thaw cycles, which can cause protein aggregation and loss of activity. Aliquot the protein upon receipt to minimize freeze-thaw events.
• Reconstitution: If supplied in lyophilized form, reconstitute the protein in the buffer recommended by the manufacturer. Centrifuge briefly after reconstitution to collect any precipitate. Allow the protein to equilibrate at room temperature before use, but do not warm above 37°C.
• Carrier Protein: For dilute working solutions, addition of a carrier protein such as BSA (0.1–0.5%) can help prevent protein adsorption to tube surfaces and maintain protein stability.
• Endotoxin Considerations: For cell-based functional assays, verify that endotoxin levels are within acceptable limits to avoid confounding biological effects.
• Experimental Controls: Always include appropriate positive and negative controls. Using the recombinant protein as a positive control in immunodetection assays helps validate assay performance across experiments.

Consult the product-specific datasheet supplied with each lot of rp366300 for definitive storage, formulation, and usage instructions.

Frequently asked questions

What is GOLPH4 and why is it important in cell biology research?

GOLPH4 (Golgi Phosphoprotein 4) is a type II transmembrane protein that resides in the Golgi apparatus and is thought to play roles in maintaining Golgi structure, membrane trafficking, and protein glycosylation. Its study is relevant to understanding fundamental secretory pathway biology and diseases linked to Golgi dysfunction, such as certain cancers and glycosylation disorders.

What expression system is used to produce Recombinant Human GOLPH4 Protein?

The specific expression system used for rp366300 should be confirmed in the product datasheet. In general, Golgi-resident transmembrane proteins like GOLPH4 may be expressed in mammalian or insect cell systems to support appropriate folding and post-translational modifications. Please refer to the lot-specific certificate of analysis for confirmed details.

How should I store Recombinant Human GOLPH4 Protein to maintain its stability?

Recombinant proteins should generally be stored at –20°C or –80°C, aliquoted to avoid repeated freeze-thaw cycles, which can degrade protein integrity. If a carrier protein such as BSA is not already included in the formulation, it may be added for stabilization at low concentrations. Always refer to the product-specific datasheet for the recommended storage buffer and conditions for rp366300.

Can Recombinant Human GOLPH4 Protein be used as a positive control in Western blotting?

Yes, high-purity recombinant GOLPH4 protein is well-suited for use as a positive control in Western blotting to confirm anti-GOLPH4 antibody specificity and detection sensitivity. Loading a known quantity of the recombinant protein alongside cell or tissue lysates helps validate that the antibody is detecting the correct molecular weight band.

What quality control tests are typically performed on this recombinant protein?

Standard quality control for recombinant proteins includes SDS-PAGE purity analysis, molecular weight verification, endotoxin level testing, and in some cases functional validation assays. Specific QC data for rp366300 are provided in the lot certificate of analysis available from Aladdin Scientific upon request.

References

1. Nakamura N. Emerging new roles of GM130, a cis-Golgi matrix protein, in higher order cell functions. J Pharmacol Sci. 2010;112(3):255-264.
2. Glick BS, Nakano A. Membrane traffic within the Golgi apparatus. Annu Rev Cell Dev Biol. 2009;25:113-132.
3. Shorter J, Warren G. Golgi architecture and inheritance. Annu Rev Cell Dev Biol. 2002;18:379-420.

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