Recombinant Human GOLGA6L1 Protein: Production & Usage Protocol

Recombinant Human GOLGA6L1 Protein is a research-grade recombinant protein derived from the human GOLGA6L1 gene, supplied by Aladdin Scientific to support studies of Golgi apparatus structure, membrane trafficking, and related cellular processes.

Product Name

Recombinant Human GOLGA6L1 Protein

SKU

rp377455

Protein Family

Golgin / Golgi autoantigen family (GOLGA6 subfamily)

Species Origin

Human (Homo sapiens)

Application

Biochemical assays, protein interaction studies, antibody validation, proteomics

Storage Recommendation

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

Background and Biological Significance

GOLGA6L1 (Golgin A6 Family Like 1) is a member of the golgin superfamily, a group of coiled-coil proteins predominantly localized to the Golgi apparatus. Golgins are known to play essential roles in maintaining Golgi structure, regulating vesicular transport, and facilitating membrane tethering events. The GOLGA6 subfamily, to which GOLGA6L1 belongs, has been identified in the context of Golgi membrane organization and intracellular trafficking pathways.

Research into GOLGA6L1 and related golgins contributes to our understanding of how the Golgi apparatus maintains its characteristic ribbon-like morphology and orchestrates the sorting and delivery of proteins and lipids throughout the cell. Dysregulation of Golgi structure and function has been implicated in a range of pathological conditions, including neurodegenerative diseases and cancer.

Reagents, Equipment, and Safety

Required Reagents:

• Recombinant Human GOLGA6L1 Protein (Aladdin Scientific, SKU: rp377455)
• Sterile deionized water or manufacturer-recommended reconstitution buffer
• Phosphate-buffered saline (PBS), pH 7.4
• Carrier protein (e.g., 0.1% BSA) for long-term stability, if applicable
• Glycerol (for preparation of working aliquots, if required)

Required Equipment:

• Microcentrifuge
• Calibrated micropipettes and sterile tips
• -80 °C ultralow freezer
• Ice bucket
• Vortex mixer (low speed)

Safety: Handle all recombinant proteins following standard laboratory biosafety guidelines. Wear appropriate PPE including gloves and lab coat. Consult the Safety Data Sheet (SDS) for this product before use.

Step-by-Step Reconstitution and Handling Protocol

Step 1 – Thawing: Remove the vial of Recombinant Human GOLGA6L1 Protein (rp377455) from -80 °C storage. Allow the vial to thaw slowly on ice or at 4 °C. Do not thaw at room temperature or use a water bath, as rapid temperature changes may reduce protein activity.

Step 2 – Brief Centrifugation: Once thawed, briefly centrifuge the vial (e.g., 500 × g for 5–10 seconds) to collect any material from the cap or walls of the tube.

Step 3 – Reconstitution: If the product is supplied in lyophilized form, reconstitute by adding the recommended volume of sterile deionized water or supplied reconstitution buffer. Refer to the product datasheet for the specific volume. Gently mix by pipetting up and down or by low-speed vortexing; do not shake vigorously.

Step 4 – Aliquoting: Prepare single-use aliquots to minimize freeze-thaw cycles. Transfer aliquots into sterile, low-binding microcentrifuge tubes. Label clearly with date and concentration.

Step 5 – Storage: Store aliquots at -80 °C. For short-term use (up to 1 week), aliquots may be kept at -20 °C. Avoid repeated freeze-thaw cycles, which can cause protein aggregation and loss of activity.

Step 6 – Working Concentration: Dilute to the desired working concentration in an appropriate assay buffer immediately before use. Determine the optimal concentration empirically for your specific assay.

Quality Control and Troubleshooting

Quality Control: Recombinant Human GOLGA6L1 Protein from Aladdin Scientific is manufactured under controlled conditions. Each lot is typically verified for purity by SDS-PAGE and/or Western blot, and for identity by mass spectrometry or equivalent methods. Refer to the Certificate of Analysis (CoA) included with each shipment for lot-specific QC data.

Common Issues and Troubleshooting:

• Protein aggregation: If visible aggregation is observed after reconstitution, centrifuge at low speed (e.g., 10,000 × g for 10 minutes at 4 °C) and use the supernatant. Ensure reconstitution buffer and conditions are as specified in the datasheet.
• Low activity or signal: Verify that the protein has not undergone excessive freeze-thaw cycles. Check that assay conditions (pH, salt concentration, temperature) are appropriate for this protein. Titrate the protein concentration to identify the optimal working range.
• Unexpected bands on SDS-PAGE: Minor bands may represent protein isoforms, degradation products, or glycosylation variants. Refer to the CoA for expected band patterns.

Recommended Applications and Citations

Recombinant Human GOLGA6L1 Protein is suitable for a range of research applications, including but not limited to:

• Protein–protein interaction studies: Use as bait or prey in pull-down assays, co-immunoprecipitation, or yeast two-hybrid validation experiments to map GOLGA6L1 binding partners.
• Antibody validation: Serve as a positive control antigen for ELISA, dot blot, or Western blot to confirm antibody specificity against GOLGA6L1.
• Biochemical characterization: Investigate coiled-coil domain function, binding kinetics, or post-translational modifications using purified recombinant protein.
• Proteomics and structural studies: Use as a standard or reference material in mass spectrometry-based proteomics or structural biology experiments.

When publishing data generated using this product, please cite the product name (Recombinant Human GOLGA6L1 Protein) and catalog number (rp377455), and Aladdin Scientific as the supplier, in accordance with journal guidelines.

Frequently asked questions

How should Recombinant Human GOLGA6L1 Protein be stored to maintain stability?

Store the protein at -80 °C in single-use aliquots to avoid repeated freeze-thaw cycles. For short-term use within one week, storage at -20 °C is acceptable. Adding a carrier protein such as 0.1% BSA can help stabilize the protein during long-term storage.

What is the biological function of GOLGA6L1?

GOLGA6L1 is a member of the golgin superfamily of coiled-coil proteins associated with the Golgi apparatus. Golgins are involved in maintaining Golgi structure, vesicular tethering, and intracellular trafficking. The specific functional characterization of GOLGA6L1 is an active area of research.

Which assays is Recombinant Human GOLGA6L1 Protein (rp377455) validated for?

This recombinant protein is suitable for use in protein–protein interaction assays, antibody validation (ELISA, Western blot, dot blot), biochemical characterization studies, and proteomics applications. Please refer to the product datasheet and Certificate of Analysis for lot-specific validation data.

How do I reconstitute the lyophilized form of GOLGA6L1 Protein?

Add the recommended volume of sterile deionized water or the supplier-specified reconstitution buffer to the lyophilized vial. Mix gently by low-speed pipetting or vortexing; do not shake vigorously. Allow the solution to sit briefly on ice before aliquoting and storing at -80 °C.

Can GOLGA6L1 Protein be used in cell-based assays?

While recombinant GOLGA6L1 protein is primarily intended for cell-free biochemical assays and protein interaction studies, potential cell-based applications would require additional optimization and validation by the end user to ensure appropriate delivery, activity, and non-toxicity under the specific experimental conditions.

References

1. Munro S. The golgin coiled-coil proteins of the Golgi apparatus. Cold Spring Harb Perspect Biol. 2011;3(6):a005256. doi:10.1101/cshperspect.a005256
2. Barr FA, Short B. Golgins in the structure and dynamics of the Golgi apparatus. Curr Opin Cell Biol. 2003;15(4):405-413. doi:10.1016/S0955-0674(03)00054-1
3. Aladdin Scientific. Recombinant Human GOLGA6L1 Protein, SKU: rp377455. Product Datasheet.

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