Recombinant Human GJA8 Protein: Handling & Usage Protocol

Recombinant Human GJA8 Protein is a research-grade connexin 50 protein produced for use in gap-junction biology, lens physiology studies, and cataract-associated mutation research.

Product Name

Recombinant Human GJA8 Protein

SKU

rp370641

Also Known As

Connexin 50 (Cx50), Gap Junction Alpha-8 Protein

Species

Human (Homo sapiens)

Expression System

Recombinant (expression system to be confirmed with product datasheet)

Recommended Storage

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

Application

In vitro biochemical assays, structural studies, antibody validation, gap-junction research

Background: GJA8 and Gap Junction Biology

GJA8 (Gap Junction Alpha-8 Protein), commonly designated Connexin 50 (Cx50), is a member of the connexin family of transmembrane proteins responsible for forming intercellular gap-junction channels. These channels facilitate direct cytoplasmic communication between adjacent cells by allowing the passage of ions, small metabolites, and signaling molecules up to approximately 1 kDa.

In humans, GJA8 is predominantly expressed in the ocular lens, where it plays an essential role in maintaining lens transparency and homeostasis. Pathogenic mutations in the GJA8 gene are associated with autosomal dominant congenital cataracts, making the protein a key subject of ophthalmological and structural biology research.

Recombinant Human GJA8 Protein enables researchers to investigate connexin-50 channel assembly, protein–protein interactions, and the functional consequences of disease-linked variants in a controlled in vitro setting.

Reconstitution Protocol

Follow the steps below to reconstitute Recombinant Human GJA8 Protein. Always consult the product-specific datasheet (SKU: rp370641) for exact buffer composition, concentration, and volume recommendations before proceeding.

1. Equilibrate the vial: Remove the vial from frozen storage and allow it to equilibrate at room temperature for 10–15 minutes. Do not expose the lyophilized protein to direct heat.
2. Prepare reconstitution buffer: Use sterile deionized water or the buffer specified in the product datasheet (typically PBS, pH 7.4, with carrier protein such as 0.1% BSA if required). Prepare the buffer fresh or use a validated stock.
3. Add buffer gently: Pipette the recommended volume of reconstitution buffer down the inner wall of the vial. Do not vortex; instead, gently swirl or rotate the vial to dissolve the protein completely.
4. Incubate briefly: Allow the reconstituted solution to stand at room temperature for 5–10 minutes to ensure complete dissolution.
5. Centrifuge: Briefly centrifuge (300–500 × g, 1–2 minutes) to collect the solution at the bottom of the vial and remove any insoluble particulates.
6. Aliquot and store: Divide into single-use aliquots to minimize freeze-thaw cycles and store as directed (see Storage section).

Note: Exact buffer composition and reconstitution volume must be confirmed from the official product datasheet for rp370641.

Storage and Stability

Proper storage is critical to maintaining the activity and integrity of Recombinant Human GJA8 Protein.

• Long-term storage: Store lyophilized protein at -20°C or -80°C. Lyophilized preparations are generally stable for 12 months or longer when stored correctly; confirm exact shelf-life from the datasheet.
• Reconstituted protein: Once reconstituted, use immediately or aliquot and store at -80°C. Reconstituted protein may be stored at 4°C for short-term use (typically up to 1 week) if a carrier protein such as BSA has been added; confirm with the datasheet.
• Avoid repeated freeze-thaw cycles: Each freeze-thaw cycle can lead to protein aggregation and loss of activity. Single-use aliquots are strongly recommended.
• Carrier protein: Adding 0.1% BSA or human serum albumin to the reconstitution buffer can help stabilize dilute protein solutions and reduce non-specific adsorption to tube surfaces.

All storage conditions should be verified against the official rp370641 product datasheet.

Experimental Applications

Recombinant Human GJA8 Protein (rp370641) is suitable for a range of in vitro research applications, including but not limited to:

• Biochemical assays: Use as a positive control or standard in ELISA, Western blotting, and dot-blot assays designed to detect or quantify GJA8/Connexin 50.
• Antibody validation: Suitable for confirming the specificity of anti-GJA8 antibodies in immunoassay development.
• Protein–protein interaction studies: Can be used in pull-down assays, co-immunoprecipitation setups, or surface plasmon resonance experiments to characterize connexin-50 binding partners.
• Structural and biophysical studies: Supports investigations into connexin oligomerization, hemichannel and full channel assembly in vitro.
• Disease mutation research: Provides a wild-type reference for comparative studies on cataract-associated GJA8 variants.

Users should optimize assay conditions, protein concentrations, and buffer systems for each specific application. Refer to the product datasheet for validated application data.

Quality, Safety, and Compliance Notes

Quality: Recombinant Human GJA8 Protein from Aladdin Scientific is manufactured under controlled conditions. Purity, endotoxin levels, and biological activity are assessed per lot; refer to the Certificate of Analysis (CoA) accompanying your specific lot for precise specifications.

Safety: This product is intended for research use only (RUO) and is not approved for diagnostic or therapeutic use in humans or animals. Handle in accordance with good laboratory practice (GLP). Consult the Safety Data Sheet (SDS) for hazard information, PPE requirements, and disposal guidelines.

Regulatory: Not for clinical, veterinary, or food-processing use. Users are responsible for ensuring compliance with applicable local regulations when using or disposing of this product.

Lot-to-lot consistency: Always record the lot number from the vial label. If performance differs unexpectedly between lots, contact Aladdin Scientific technical support and retain the CoA for reference.

Frequently asked questions

What is GJA8 protein and why is it important in research?

GJA8 encodes Connexin 50 (Cx50), a gap-junction protein critical for intercellular communication in the ocular lens. Mutations in GJA8 are linked to autosomal dominant congenital cataracts, making it a primary target for understanding lens biology and developing insights into connexin-related diseases.

How should I reconstitute and store Recombinant Human GJA8 Protein to preserve activity?

Reconstitute the lyophilized protein using sterile PBS (pH 7.4) or the buffer specified in the product datasheet. Avoid vortexing; gently swirl to dissolve. Aliquot the reconstituted protein into single-use volumes and store at -80°C. Avoid repeated freeze-thaw cycles, as these reduce protein activity and promote aggregation. Always confirm exact conditions with the rp370641 datasheet.

What experimental applications is rp370641 validated for?

Recombinant Human GJA8 Protein (rp370641) is suitable for in vitro applications including ELISA, Western blotting, antibody specificity validation, protein–protein interaction studies (e.g., pull-down, SPR), and as a wild-type reference in cataract mutation research. Application-specific validated data should be confirmed in the official product datasheet.

Can Recombinant Human GJA8 Protein be used in cell-based assays?

Recombinant Human GJA8 Protein is primarily intended for cell-free, in vitro biochemical and biophysical applications. Its suitability for cell-based functional assays depends on purity, endotoxin levels, and formulation. Consult the product datasheet and Certificate of Analysis for lot-specific endotoxin data before use in cell culture systems.

What should I do if the protein does not dissolve completely after reconstitution?

If particulates remain after gentle swirling, briefly centrifuge the vial at 300–500 × g for 1–2 minutes and transfer the supernatant. If the problem persists, try warming the solution briefly at 37°C for no more than 5 minutes while gently mixing. Contact Aladdin Scientific technical support if complete dissolution cannot be achieved, and retain the lot number and CoA for troubleshooting.

References

1. White TW, Bruzzone R. Multiple connexin proteins in single intercellular channels: connexin compatibility and functional consequences. J Bioenerg Biomembr. 1996;28(4):339-350.
2. Shiels A, Bhatt DL, Bhatt P. Connexin 50 and connexin 46: in the lens you are connected. Trends Mol Med. 2007; (general reference – specific details to be verified).
3. Aladdin Scientific. Recombinant Human GJA8 Protein, SKU rp370641. Product Datasheet. Aladdin Scientific; [date to be confirmed].

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