Recombinant Human GJD2 Protein: Research Guide & Applications

Recombinant Human GJD2 Protein is a recombinantly expressed form of the gap junction delta-2 protein (Connexin-36), offering researchers a well-defined reagent for studying electrical synaptic transmission, neuronal coupling, and connexin biology.

Product Name

Recombinant Human GJD2 Protein

SKU

rp369257

Gene Name

GJD2 (also known as CX36, CONNEXIN-36)

Protein Family

Connexin (Gap Junction) protein family

Primary Research Area

Neuroscience; electrical synapse; gap junction biology

Source Reference

absea #PP-12657

Biological Background of GJD2 (Connexin-36)

GJD2, commonly referred to as Connexin-36 (Cx36), is a member of the connexin superfamily of proteins responsible for forming gap junctions — specialized intercellular channels that enable direct cytoplasmic communication between adjacent cells. Unlike many connexins that are broadly expressed, Cx36 exhibits a highly restricted expression pattern, being predominantly found in neurons of the central nervous system (CNS), including interneurons of the cerebral cortex, hippocampus, retina, and inferior olive.

At the molecular level, Cx36 assembles into hexameric hemichannels (connexons) that dock with complementary connexons on adjacent cells to form complete gap junction channels. These channels are permeable to small molecules, ions, and second messengers (typically <1 kDa), thereby mediating electrical coupling and metabolic coordination between neurons.

Functionally, GJD2-mediated gap junctions underpin electrical synaptic transmission, neural synchrony, and oscillatory network activity. Research has implicated Cx36 in processes such as visual processing in the retina, gamma oscillations in the neocortex, insulin secretion in pancreatic beta cells, and the modulation of seizure activity. Loss-of-function studies in rodent models have demonstrated its importance in high-frequency neuronal synchronization.

Protein Characteristics and Formulation Guidelines

Recombinant Human GJD2 Protein (rp369257) is produced using recombinant expression technology to deliver a consistent, research-grade reagent. When working with this protein, researchers should note the following general considerations applicable to recombinant membrane-associated or full-length connexin proteins:

• Purity: Recombinant preparations are typically subjected to affinity chromatography and quality-control analysis to confirm identity and purity. Researchers should verify the certificate of analysis (CoA) supplied with each lot for specific purity data.
• Formulation: Recombinant connexin proteins are often supplied in a buffered solution containing stabilizing agents. Consult the product datasheet for exact buffer composition, pH, and excipient details for lot rp369257.
• Molecular Weight: The predicted molecular weight of human Cx36 (GJD2) is approximately 36 kDa, though apparent molecular weight on SDS-PAGE may differ due to post-translational characteristics of the recombinant system used.
• Endotoxin Levels: For cell-based assays, confirm endotoxin levels from the CoA to ensure results are not confounded by LPS contamination.

Always centrifuge the vial briefly before opening and avoid repeated freeze-thaw cycles, which may compromise protein integrity and activity.

Storage, Handling, and Reconstitution Protocol

Proper storage and handling are critical for maintaining the biological activity and structural integrity of Recombinant Human GJD2 Protein. Follow these guidelines:

1. Long-term Storage: Store the protein at −80 °C upon receipt if it will not be used within 1–2 weeks. Avoid storing at −20 °C for extended periods as repeated temperature cycling can cause aggregation.
2. Short-term Storage: For working aliquots intended for use within one week, storage at 4 °C in a stable buffer is generally acceptable. Minimize exposure to ambient temperature.
3. Aliquoting: Upon first thaw, prepare single-use aliquots to avoid repeated freeze-thaw cycles. Label aliquots with date and lot number for traceability.
4. Reconstitution (if lyophilized): If supplied in lyophilized form, reconstitute by adding the volume of sterile, deionized water or the recommended buffer as specified on the product datasheet. Allow the vial to stand at room temperature for 10 minutes, then gently mix by pipetting. Do not vortex.
5. Working Concentration: Determine optimal working concentration empirically for each assay type. Pilot experiments using a concentration range are recommended before large-scale studies.
6. Carrier Protein: Addition of a carrier protein (e.g., 0.1% BSA) to dilution buffers may help maintain protein stability at low concentrations.

Always consult the current product datasheet for lot-specific storage instructions, as formulation details may vary between lots.

Research Applications of Recombinant Human GJD2 Protein

Recombinant Human GJD2 Protein (rp369257) is suitable for a range of research applications in neuroscience, cell biology, and pharmacology. Principal applications include:

• Protein–Protein Interaction Studies: GJD2 recombinant protein can be used in pull-down assays, co-immunoprecipitation validation experiments, and surface plasmon resonance (SPR) studies to characterize Cx36 interactome partners, including scaffolding proteins and regulatory kinases.
• Antibody Validation: As a positive control antigen, the recombinant protein is used to validate anti-GJD2/Cx36 antibodies in Western blot, ELISA, and immunofluorescence applications, ensuring antibody specificity prior to use in complex biological samples.
• Biochemical Assays: The protein can be employed in enzyme-linked immunosorbent assays (ELISAs) for measuring Cx36 levels or in competitive binding assays to investigate small-molecule modulators of gap junction assembly.
• Structural Studies: Recombinant Cx36 preparations serve as material for biophysical characterization techniques such as circular dichroism (CD) spectroscopy or dynamic light scattering (DLS) to assess secondary structure and aggregation state.
• Drug Discovery Screening: Cx36 is an emerging target in epilepsy, retinal degeneration, and diabetes research. Recombinant GJD2 protein facilitates high-throughput screening of chemical libraries for modulators of Cx36 function.

Researchers should optimize assay conditions specific to their experimental system and refer to published literature for guidance on established GJD2 assay formats.

Quality Assurance and Ordering Information

Aladdin Scientific is committed to providing research-grade reagents supported by rigorous quality control. For Recombinant Human GJD2 Protein (rp369257), the following quality and ordering details apply:

• SKU: rp369257
• Reference: absea #PP-12657
• Quality Control: Each lot is tested for purity, identity (e.g., by SDS-PAGE and/or Western blot), and biological activity where applicable. A Certificate of Analysis (CoA) is available upon request for each specific lot number.
• Safety Data Sheet (SDS): An SDS is available for download from the Aladdin Scientific website. Consult the SDS for safe handling, disposal, and first-aid procedures.
• Custom Requirements: For bulk quantities, custom formulations, or special packaging, contact Aladdin Scientific technical support or your regional sales representative.
• Returns and Replacements: Damaged or non-conforming products should be reported to customer service within the stipulated timeframe as detailed in Aladdin Scientific's returns and refunds policy.

To place an order or request technical support for rp369257, visit the Aladdin Scientific website or contact your local distributor.

Frequently asked questions

What is GJD2 protein and why is it important in neuroscience research?

GJD2, also known as Connexin-36 (Cx36), is a gap junction protein predominantly expressed in neurons of the central nervous system. It forms intercellular channels that mediate electrical synaptic transmission and neural synchrony. Its importance stems from roles in gamma oscillations, retinal signal processing, pancreatic beta-cell coupling, and modulation of seizure susceptibility, making it a key target in neuroscience and metabolic disease research.

How should Recombinant Human GJD2 Protein (rp369257) be stored to maintain activity?

For long-term storage, keep the protein at −80 °C. If the protein will be used within one week, it may be stored at 4 °C. Avoid repeated freeze-thaw cycles by preparing single-use aliquots upon first thaw. Always refer to the lot-specific product datasheet and Certificate of Analysis for the most accurate storage recommendations.

What assay types is Recombinant Human GJD2 Protein suitable for?

Recombinant Human GJD2 Protein (rp369257) is suitable for multiple applications including antibody validation (Western blot, ELISA, immunofluorescence), protein-protein interaction studies (pull-down, SPR), biochemical and binding assays, structural characterization (CD spectroscopy, DLS), and drug discovery screening. Researchers should empirically optimize conditions for each specific application.

How can I request a Certificate of Analysis (CoA) for rp369257?

A Certificate of Analysis for Recombinant Human GJD2 Protein (rp369257) can be requested through the Aladdin Scientific website or by contacting customer service with your lot number. The CoA provides lot-specific data on purity, identity, and quality control testing results.

References

1. Connors BW, Long MA. Electrical synapses in the mammalian brain. Annu Rev Neurosci. 2004;27:393-418. doi:10.1146/annurev.neuro.26.041002.131128
2. Söhl G, Maxeiner S, Willecke K. Expression and functions of neuronal gap junctions. Nat Rev Neurosci. 2005;6(3):191-200. doi:10.1038/nrn1627
3. Deans MR, Gibson JR, Bhatt DL, et al. Synchronous activity of inhibitory networks in neocortex requires electrical synapses containing connexin36. Neuron. 2001;31(3):477-485. doi:10.1016/S0896-6273(01)00373-7

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