Recombinant Human GCC1 Protein: Properties & Research Guide

Recombinant Human GCC1 Protein is a research-grade recombinant protein derived from the human GCC1 gene, designed to support investigations into Golgi membrane trafficking, vesicle tethering, and related intracellular transport mechanisms.

Product Name

Recombinant Human GCC1 Protein

SKU

rp374299

Gene / Protein Target

GCC1 (GRIP and coiled-coil domain-containing protein 1, also known as GCC88)

Species Origin

Human (Homo sapiens)

Protein Function

Peripheral membrane protein involved in vesicle tethering at the trans-Golgi network (TGN) and endosomal trafficking

Recommended Storage

Store at -80°C; avoid repeated freeze-thaw cycles

Primary Applications

ELISA, Western blot, functional assays, protein interaction studies

Biological Background of GCC1

GCC1, also referred to as GCC88, is a peripheral membrane protein that localizes predominantly to the trans-Golgi network (TGN). It belongs to the golgin family of coiled-coil tethering factors, which play essential roles in maintaining Golgi architecture and facilitating membrane trafficking between the Golgi apparatus and endosomal compartments.

The protein contains a C-terminal GRIP domain responsible for TGN localization, and extended coiled-coil regions that mediate vesicle capture and tethering. GCC1 has been implicated in retrograde transport pathways, specifically in the retrieval of cargo from early and recycling endosomes back to the TGN. Its interaction with small GTPases of the Rab and Arl families is critical for membrane recruitment and functional activity.

Research into GCC1 is relevant to understanding fundamental cell biology processes including protein sorting, membrane identity, and intracellular trafficking, as well as pathological conditions where Golgi integrity or endosomal trafficking is disrupted.

Protein Specifications and Quality Attributes

Aladdin Scientific's Recombinant Human GCC1 Protein (rp374299) is produced using recombinant expression systems to ensure high purity and batch-to-batch consistency. Key quality attributes researchers should verify upon receipt include:

• Purity: Confirm purity level via SDS-PAGE or HPLC analysis as indicated on the Certificate of Analysis (CoA). High-purity preparations minimize background signal in sensitive assays.
• Endotoxin Level: Low endotoxin content is critical for cell-based functional assays; refer to the product CoA for the tested endotoxin specification.
• Formulation: The protein is typically supplied in a buffered solution or lyophilized format. Confirm the exact formulation on the product datasheet before experimental use.
• Molecular Weight: Verify observed molecular weight against the theoretical value by Western blot, accounting for any tags or post-translational modifications present in the recombinant construct.

Always request and review the CoA for each lot to confirm that release criteria have been met prior to integrating the protein into critical assays.

Reconstitution, Handling, and Storage Guidelines

Proper handling of Recombinant Human GCC1 Protein is essential to preserve its structural integrity and biological activity. Follow these guidelines:

Reconstitution (if lyophilized)

• Briefly centrifuge the vial before opening to collect lyophilized material at the bottom.
• Reconstitute with the volume of sterile, ultrapure water or appropriate buffer as specified on the product datasheet to achieve the recommended concentration.
• Gently mix by pipetting; do not vortex, as shear forces can denature the protein.
• Allow the reconstituted solution to equilibrate at room temperature for 5–10 minutes before use.

Storage

• Store the reconstituted or liquid protein at -80°C for long-term storage.
• For short-term use (up to 1 week), aliquots may be stored at -20°C or 4°C as specified in the datasheet.
• Prepare single-use aliquots to avoid repeated freeze-thaw cycles, which can cause protein aggregation and loss of activity.

General Handling Tips

• Work on ice and use pre-chilled tubes and pipette tips when handling the protein.
• Avoid prolonged exposure to room temperature.
• Use protein low-binding tubes to minimize adsorptive losses.

Recommended Applications and Experimental Considerations

Recombinant Human GCC1 Protein is suitable for a range of biochemical and cell biological research applications. Researchers should conduct empirical optimization for each specific assay format:

• ELISA: Use as a standard or coating antigen for development and validation of GCC1-specific immunoassays. Prepare serial dilutions in assay buffer to establish a standard curve.
• Western Blot (WB): Employ as a positive control to confirm antibody specificity and optimize detection conditions. Load appropriate amounts alongside cell or tissue lysates.
• Protein–Protein Interaction Studies: Use in pull-down assays, co-immunoprecipitation validation, or surface plasmon resonance (SPR) experiments to characterize interactions between GCC1 and candidate binding partners such as Rab GTPases or GRIP-domain-interacting proteins.
• Functional / Biochemical Assays: Explore GCC1 tethering activity in in vitro reconstitution systems or test its role in modulating vesicle fusion kinetics.
• Structural Studies: High-purity recombinant protein can be used as input material for crystallography or cryo-EM studies aimed at resolving GCC1 domain architecture.

Always include appropriate positive and negative controls. Optimize protein concentration, incubation time, and buffer conditions empirically for each application.

Frequently asked questions

What is the biological function of GCC1 and why is it relevant to my research?

GCC1 (GCC88) is a golgin family tethering factor that localizes to the trans-Golgi network and participates in endosome-to-TGN retrograde trafficking. It is relevant to research in membrane trafficking, Golgi biology, protein sorting, and diseases involving intracellular transport defects. Recombinant GCC1 protein enables biochemical dissection of these pathways in vitro.

How should I reconstitute the lyophilized Recombinant Human GCC1 Protein?

Briefly centrifuge the vial to collect the lyophilized material, then add sterile ultrapure water or the buffer recommended on the product datasheet to achieve the target concentration. Mix gently by pipetting — do not vortex — and allow the solution to equilibrate at room temperature for 5–10 minutes. Prepare single-use aliquots and store at -80°C to avoid activity loss from freeze-thaw cycling.

What quality control tests are performed on Aladdin Scientific's Recombinant Human GCC1 Protein?

Quality control specifications, including purity assessment (e.g., SDS-PAGE), endotoxin testing, and biological activity data, are detailed on the Certificate of Analysis (CoA) supplied with each lot. Researchers are advised to request the lot-specific CoA from Aladdin Scientific and review all release criteria before use in critical experiments.

Can Recombinant Human GCC1 Protein be used in cell-based functional assays?

Yes, provided the endotoxin level is sufficiently low as confirmed by the CoA. For cell-based applications, use endotoxin-tested lots and optimize the protein concentration empirically. Consider the need for a delivery reagent if intracellular activity is required, and always include appropriate vehicle and negative controls.

What are the recommended storage conditions to maintain GCC1 protein activity?

Store the protein at -80°C for long-term stability and prepare single-use aliquots to prevent repeated freeze-thaw cycles. Short-term storage at -20°C or 4°C may be acceptable as specified in the product datasheet. Avoid prolonged exposure to ambient temperature and use protein low-binding tubes to minimize adsorptive losses.

References

1. Munro S, Nichols BJ. The GRIP domain - a novel Golgi-targeting domain found in several coiled-coil proteins. Curr Biol. 1999;9(7):377-380. doi:10.1016/S0960-9822(99)80166-3
2. Luke MR, Kjer-Nielsen L, Brown DL, Stow JL, Gleeson PA. GRIP domain-mediated targeting of two new coiled-coil proteins, GCC88 and GCC185, to subcompartments of the trans-Golgi network. J Biol Chem. 2003;278(6):4216-4226. doi:10.1074/jbc.M210433200
3. Aladdin Scientific. Recombinant Human GCC1 Protein (rp374299) Product Datasheet. Aladdin Scientific; [date not specified].

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