Recombinant Human GOLGA8R Protein: Preparation & Usage Protocol

Recombinant Human GOLGA8R Protein is a research-grade reagent designed for investigators studying Golgi apparatus organization, membrane trafficking, and related cellular processes requiring a well-characterized human GOLGA8R antigen.

Product Name

Recombinant Human GOLGA8R Protein

SKU

rp377485

Protein Family

Golgin family (Golgi autoantigen, golgin subfamily A)

Expression System

Recombinant (system to be confirmed with datasheet)

Recommended Storage

-20°C or -80°C; avoid repeated freeze-thaw cycles

Application

Suitable for use as antigen, ELISA standard, and in vitro functional studies (confirm with datasheet)

Materials and Equipment Required

Before beginning, ensure the following materials are available:

• Recombinant Human GOLGA8R Protein (rp377485, Aladdin Scientific)
• Sterile, ultrapure water or the reconstitution buffer specified on the product datasheet
• Microcentrifuge tubes (1.5 mL, low-binding recommended)
• Calibrated micropipettes and sterile filtered tips
• Refrigerated microcentrifuge
• Ice bucket or cooling block
• Appropriate assay buffers for downstream applications (e.g., PBS, ELISA coating buffer)
• Personal protective equipment (gloves, lab coat, eye protection)

Note: Always refer to the lot-specific Certificate of Analysis (CoA) and product datasheet supplied with the product for confirmed specifications including host expression system, purity, and formulation buffer.

Reconstitution Protocol

Follow these steps to reconstitute lyophilized Recombinant Human GOLGA8R Protein:

1. Equilibrate: Allow the vial to equilibrate to room temperature for 10–15 minutes before opening to prevent condensation-related contamination.
2. Centrifuge: Briefly centrifuge the vial (1,000–2,000 × g, 30 seconds) to pellet any lyophilized material from the walls and cap.
3. Add Reconstitution Buffer: Add the volume of sterile reconstitution buffer (as specified on the datasheet) slowly down the side of the vial. Do not pipette directly onto the protein cake.
4. Gently Mix: Swirl or gently tap the vial to dissolve. Do NOT vortex, as vigorous agitation can denature the protein.
5. Incubate: Allow the solution to sit at room temperature for 5–10 minutes, or at 4°C for up to 30 minutes, until fully dissolved.
6. Inspect: The solution should be clear. Discard if particulate matter or turbidity is observed.
7. Aliquot: Divide the reconstituted protein into single-use aliquots to minimize freeze-thaw cycles and store immediately under the recommended conditions.

Important: Confirm the exact recommended reconstitution buffer and final concentration from the lot-specific datasheet provided with the product.

Storage and Stability

Proper storage is critical to maintaining the activity and integrity of Recombinant Human GOLGA8R Protein.

• Short-term storage (≤1 week): Store at 4°C. Keep the vial tightly sealed to avoid contamination and evaporation.
• Long-term storage: Store aliquots at -20°C or -80°C. Storage at -80°C is recommended for extended periods to maximize stability.
• Freeze-thaw cycles: Limit to a maximum of 2–3 cycles. Each freeze-thaw event can progressively reduce protein activity and increase aggregation.
• Carrier protein: If the protein is supplied without a stabilizing carrier (e.g., BSA), consider adding a carrier protein or glycerol (to a final concentration of 10–50%) for long-term storage, as appropriate and confirmed with the datasheet.
• Expiry: Refer to the vial label and CoA for the stated shelf life. Do not use beyond the expiry date.

Note: Specific stability data for this product should be verified from the product datasheet. The guidelines above represent standard best practices for recombinant proteins.

Experimental Application Guidelines

Recombinant Human GOLGA8R Protein is intended for in vitro research use only. Potential applications include, but are not limited to:

• Antigen for antibody production: The protein may be used to immunize hosts for the generation of anti-GOLGA8R antibodies or as a positive control antigen in immunoassays.
• ELISA standard or coating antigen: Suitable for use as a standard or coating antigen in enzyme-linked immunosorbent assays to detect or quantify GOLGA8R-specific antibodies.
• In vitro binding assays: Can be employed to study protein–protein interactions involving GOLGA8R and its binding partners within the Golgi complex.
• Western blot positive control: A known quantity of the recombinant protein can serve as a positive control to validate anti-GOLGA8R antibody performance.

Optimize working concentrations empirically for each specific assay system. Always include appropriate negative and positive controls. Confirm suitability for each application from the product datasheet before use.

Safety and Quality Considerations

Safety: Although Recombinant Human GOLGA8R Protein is not classified as a hazardous substance under standard laboratory reagent guidelines, standard biosafety precautions should be followed:

• Wear appropriate PPE at all times: gloves, lab coat, and eye protection.
• Avoid inhalation or ingestion. Do not pipette by mouth.
• Dispose of all biological materials and contaminated consumables in accordance with institutional and local regulatory requirements.

Quality Control: Each lot of Recombinant Human GOLGA8R Protein is subject to quality control testing. Refer to the Certificate of Analysis (CoA) provided with the product for lot-specific data on purity, concentration, and endotoxin levels. Key quality attributes to verify include:

• Purity (typically assessed by SDS-PAGE or HPLC)
• Identity (confirmed by Western blot or mass spectrometry)
• Endotoxin content (particularly important for cell-based assays)
• Biological activity (where applicable, confirmed per datasheet)

Contact Aladdin Scientific technical support if your CoA or datasheet is missing, or if you observe unexpected results.

Frequently asked questions

What is GOLGA8R and why is it studied in research?

GOLGA8R (Golgi Autoantigen, Golgin Subfamily A Member 8R) is a member of the golgin family of coiled-coil proteins associated with the Golgi apparatus, which plays roles in membrane trafficking and Golgi structure maintenance. It is studied in the context of Golgi biology, intracellular transport, and as an autoantigen in certain autoimmune conditions. Confirm specific functional details from current peer-reviewed literature.

How should I reconstitute the Recombinant Human GOLGA8R Protein if no datasheet is available?

If you have not received a datasheet, contact Aladdin Scientific customer support immediately to obtain the lot-specific document. As a general best practice for lyophilized recombinant proteins, sterile ultrapure water or PBS (pH 7.4) is commonly used for reconstitution; however, the specific buffer and volume must be confirmed from the official datasheet to avoid compromising protein integrity.

Can Recombinant Human GOLGA8R Protein be used in cell-based assays?

Potential suitability for cell-based assays depends on the endotoxin level, formulation, and confirmed biological activity of the specific lot. Always check the Certificate of Analysis for endotoxin content before using in cell-based systems, as elevated endotoxin can confound results. Confirm application suitability with the product datasheet or Aladdin Scientific technical support.

How many freeze-thaw cycles can the protein withstand?

As a general guideline for recombinant proteins, it is recommended to limit freeze-thaw cycles to no more than 2–3 to preserve activity and minimize aggregation. Aliquoting the protein upon reconstitution into single-use volumes is the best strategy to avoid repeated freeze-thaw. Verify specific stability data from the product datasheet.

References

1. Aladdin Scientific. Recombinant Human GOLGA8R Protein, SKU rp377485. Product datasheet and Certificate of Analysis.
2. Shorter J, Warren G. Golgi architecture and inheritance. Annual Review of Cell and Developmental Biology. 2002;18:379–420.
3. Munro S. The golgin coiled-coil proteins of the Golgi apparatus. Cold Spring Harbor Perspectives in Biology. 2011;3(6):a005256.

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