Recombinant Human GOLGA8K Protein: Handling & Usage Protocol
Recombinant Human GOLGA8K Protein: Handling & Usage Protocol
Recombinant Human GOLGA8K Protein is a research-grade reagent targeting the GOLGA8K golgin-family member, requiring careful reconstitution, precise aliquoting, and controlled storage to maintain full biological activity for downstream applications.
- Product Name
- Recombinant Human GOLGA8K Protein
- SKU
- rp377478
- Protein Family
- Golgin family (Golgi apparatus-associated proteins)
- Host / Expression System
- Recombinant expression system (verify datasheet for specific host)
- Recommended Storage
- -20°C to -80°C; avoid repeated freeze-thaw cycles
- Application
- Suitable for use as antigen, in binding assays, and related in vitro research applications
Materials and Equipment Required
Before beginning, ensure the following materials and equipment are available:
- Recombinant Human GOLGA8K Protein (SKU: rp377478)
- Sterile ultrapure water or the reconstitution buffer specified on the product datasheet
- Microcentrifuge tubes (low-binding, 0.5 mL or 1.5 mL)
- Calibrated micropipettes and sterile tips
- Benchtop microcentrifuge
- –20°C or –80°C freezer for long-term storage
- Ice bucket or cooling block
- Protein quantification reagents (e.g., BCA or Bradford assay kit)
Note: Always consult the lot-specific Certificate of Analysis (CoA) and product datasheet supplied with SKU rp377478 for host system, purity, formulation, and concentration details prior to use, as these may vary by lot.
Reconstitution Procedure
Follow these steps to reconstitute Recombinant Human GOLGA8K Protein correctly:
- Equilibrate the vial: Allow the lyophilized protein vial to equilibrate to room temperature for 10–15 minutes before opening. Do not expose to direct sunlight or heat sources.
- Centrifuge the vial: Briefly centrifuge the sealed vial (1,000–2,000 × g, 30 seconds) to collect any residual powder from the sides and cap of the vial.
- Add reconstitution buffer: Add the volume of sterile ultrapure water or buffer recommended on the product datasheet directly to the lyophilized cake. A typical target concentration is 0.1–1.0 mg/mL; confirm the exact volume with the CoA.
- Dissolve gently: Do not vortex. Gently swirl or pipette to dissolve. Allow the solution to stand for 5 minutes at room temperature if necessary.
- Verify dissolution: Ensure the solution is clear and free of visible particulates. If cloudiness persists, centrifuge at 10,000 × g for 5 minutes at 4°C and use the supernatant.
- Quantify protein: Confirm final concentration using a BCA or Bradford assay against an appropriate standard.
Important: Specific reconstitution buffer composition, volume, and target concentration must be taken from the lot-specific datasheet for rp377478, as formulations may vary.
Storage and Stability Guidelines
Proper storage is critical to maintaining the activity and integrity of Recombinant Human GOLGA8K Protein:
- Short-term (up to 1 week): Store reconstituted protein at 4°C. Add a carrier protein such as 0.1% BSA if the protein will be used over several days to minimize adsorption to tube surfaces.
- Long-term: Aliquot into single-use volumes and store at –20°C (preferred) or –80°C for extended stability. Lyophilized protein may be stored at –20°C until reconstitution.
- Avoid freeze-thaw cycles: Repeated freeze-thaw cycles degrade protein activity. Prepare aliquots sized for single experimental use.
- Working solutions: Prepare fresh working dilutions on the day of the experiment. Do not store diluted working solutions.
- Shelf life: Refer to the expiration date on the product label and CoA. Typical lyophilized protein shelf life is 12–24 months under recommended storage conditions, but confirm with the lot-specific CoA for rp377478.
Application and Working Concentration Recommendations
Recombinant Human GOLGA8K Protein is intended for in vitro research use. Potential applications include:
- Use as an immunogen/antigen: GOLGA8K protein may be used to generate or validate antibodies targeting the GOLGA8K antigen in immunological assays.
- Binding and interaction assays: Suitable for pull-down assays, co-immunoprecipitation controls, or surface plasmon resonance (SPR) experiments investigating protein–protein interactions involving GOLGA8K.
- Functional studies: May support studies of Golgi apparatus structure, vesicular trafficking, or golgin-related cellular processes in vitro.
Working concentrations: Optimal concentrations are assay-dependent. Titrate the protein for each specific application. Begin with the manufacturer's recommended starting concentration stated on the datasheet and optimize empirically.
Compatibility: Confirm buffer compatibility with your assay system. Avoid sodium azide-containing buffers if the protein will be used in cell-based assays, as azide is cytotoxic.
All applications should be validated by the end user. This product is for research use only and is not intended for diagnostic or therapeutic use.
Troubleshooting
If you encounter issues when working with Recombinant Human GOLGA8K Protein, consult the following guidance:
| Problem | Possible Cause | Suggested Action |
|---|---|---|
| Protein will not dissolve | Insufficient reconstitution volume or incorrect buffer | Add additional buffer incrementally; ensure buffer matches datasheet recommendation |
| Cloudy or aggregated solution | Protein aggregation due to improper handling or freeze-thaw damage | Centrifuge at 10,000 × g for 5 min at 4°C; use supernatant only |
| Low or no activity in assay | Protein degradation, incorrect concentration, or incompatible assay conditions | Verify storage history; re-quantify protein; optimize assay buffer and concentration |
| High background in binding assay | Non-specific protein adsorption to surfaces | Add 0.05–0.1% BSA as carrier; use low-binding tubes and plates |
| Unexpected molecular weight on SDS-PAGE | Post-translational modifications, tags, or glycosylation | Consult CoA for expected MW; confirm reducing vs. non-reducing conditions |
For further technical support, contact Aladdin Scientific technical services with your lot number and CoA details.
Frequently asked questions
What is GOLGA8K and why is recombinant GOLGA8K protein used in research?
GOLGA8K (Golgin A8 Family Member K) is a member of the golgin family of coiled-coil proteins associated with the Golgi apparatus, involved in maintaining Golgi structure and vesicular trafficking. Recombinant Human GOLGA8K Protein is used in research to investigate these cellular functions, generate or validate specific antibodies, and study protein–protein interactions in vitro.
How should I reconstitute and store Recombinant Human GOLGA8K Protein (rp377478)?
Reconstitute the lyophilized protein using sterile ultrapure water or the buffer specified in the lot-specific datasheet, gently swirling to dissolve without vortexing. For short-term use (up to one week), store reconstituted protein at 4°C; for long-term storage, aliquot into single-use volumes and store at –20°C or –80°C. Always avoid repeated freeze-thaw cycles to preserve protein integrity.
What applications is Recombinant Human GOLGA8K Protein suitable for?
Recombinant Human GOLGA8K Protein is suitable for use as an immunogen for antibody production or validation, in protein binding and interaction assays (such as pull-down or SPR experiments), and for in vitro functional studies of Golgi-related cellular processes. It is intended for research use only and is not approved for diagnostic or therapeutic applications.
What should I do if my GOLGA8K protein solution appears cloudy after reconstitution?
If cloudiness is observed after reconstitution, centrifuge the solution at 10,000 × g for 5 minutes at 4°C and carefully collect the supernatant for use. Cloudiness may indicate protein aggregation caused by improper handling, excessive freeze-thaw cycling, or buffer incompatibility. Contact Aladdin Scientific technical support with your lot number if the problem persists.
References
- Aladdin Scientific. Recombinant Human GOLGA8K Protein (rp377478) Product Datasheet and Certificate of Analysis. Aladdin Scientific.
- Gillingham AK, Munro S. The small G proteins of the Arf family and their regulators. Annual Review of Cell and Developmental Biology. 2007;23:579-611. [General reference for golgin biology and Golgi apparatus function — specific applicability to GOLGA8K needs verification.]
- Witkos TM, Lowe M. The Golgin Family of Coiled-Coil Tethering Proteins. Frontiers in Cell and Developmental Biology. 2016;3:86. [General golgin family reference — GOLGA8K-specific data not confirmed from source material.]
Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.
危险品化学品经营许可证（带存储）