Recombinant Human GPR158 Protein: Production & Usage Protocol

Recombinant Human GPR158 Protein is a purified recombinant form of the orphan G protein-coupled receptor GPR158, produced for use in binding assays, structural studies, and GPCR-related drug discovery research.

Product Name

Recombinant Human GPR158 Protein

SKU

rp368052

Protein Family

Class C G protein-coupled receptor (GPCR), orphan receptor

Species Origin

Human (Homo sapiens)

Recommended Storage

-80°C for long-term storage; avoid repeated freeze-thaw cycles

Supplier Reference

absea#PQ-843

Overview of GPR158 and Research Significance

GPR158 is a member of the Class C G protein-coupled receptor (GPCR) superfamily and is classified as an orphan receptor, meaning its endogenous ligand(s) have not been definitively identified. It is predominantly expressed in the central nervous system, with high expression levels reported in the cerebellum, prefrontal cortex, and hippocampus. Emerging research has implicated GPR158 in mood regulation, stress responses, and potential roles in neuropsychiatric disorders such as depression. The receptor has also been identified as a candidate target for novel therapeutics due to its selective neural expression pattern.

Recombinant Human GPR158 Protein (rp368052) enables researchers to investigate receptor structure, screen for potential ligands, and perform biochemical characterization assays without reliance on endogenous expression systems. This product is intended for in vitro research use only.

Reconstitution Protocol

Follow the steps below to reconstitute Recombinant Human GPR158 Protein correctly:

1. Equilibration: Allow the vial to equilibrate to room temperature for 10–15 minutes before opening. Do not heat the vial.
2. Centrifugation: Briefly centrifuge the vial (1,000–2,000 × g, 30 seconds) to pellet any lyophilized material that may have adhered to the cap or sides of the vial.
3. Reconstitution buffer: Reconstitute the protein in sterile, deionized water or an appropriate physiological buffer (e.g., PBS, pH 7.4) to a working concentration. Refer to the Certificate of Analysis (CoA) for the supplier-recommended volume and final concentration.
4. Gentle mixing: Gently swirl or pipette to mix. Do not vortex, as this may denature the protein.
5. Carrier protein (optional): For long-term stability, consider adding a carrier protein such as 0.1% BSA (bovine serum albumin) to the reconstitution buffer, particularly when working at low protein concentrations.

Note: Always consult the product-specific CoA (absea#PQ-843) for lot-specific reconstitution instructions, as specifications may vary between lots.

Storage and Handling Guidelines

Proper storage is critical to maintaining the biological activity and structural integrity of Recombinant Human GPR158 Protein:

• Short-term storage (≤1 week): Store at 4°C after reconstitution.
• Long-term storage: Aliquot the reconstituted protein into single-use volumes and store at -80°C. This prevents degradation caused by repeated freeze-thaw cycles.
• Avoid repeated freeze-thaw cycles: Each freeze-thaw cycle can reduce protein activity. Prepare aliquots appropriate for single experiments.
• Lyophilized stock: Unopened lyophilized protein should be stored as specified on the product label and CoA, typically at -20°C or -80°C.
• Sterile technique: Handle reconstituted protein under aseptic conditions to prevent microbial contamination.

Discard any unused protein that shows signs of aggregation, turbidity, or contamination.

Recommended Experimental Applications

Recombinant Human GPR158 Protein (rp368052) is suited for the following research applications:

• Ligand binding assays: Use GPR158 protein to screen small molecule libraries or peptide collections for potential receptor ligands, supporting deorphanization efforts.
• Protein–protein interaction studies: Investigate interactions between GPR158 and intracellular signaling partners, such as G proteins or scaffolding proteins (e.g., RGS7 complex components).
• Structural biology: Suitable as an antigen for antibody generation or as a component in cryo-EM and X-ray crystallography studies aimed at resolving GPR158 structure.
• ELISA and Western blot (as antigen/standard): May be used as a protein standard or positive control in immunoassay development and validation.
• Drug discovery: Supports high-throughput screening campaigns targeting GPR158 as a novel CNS drug target.

Important: Optimize assay conditions (protein concentration, buffer composition, incubation time, and temperature) empirically for each specific application. This product is for research use only (RUO) and is not intended for diagnostic or therapeutic use.

Quality Control and Troubleshooting

Quality Control: Each lot of Recombinant Human GPR158 Protein is tested for purity and identity prior to release. Refer to the Certificate of Analysis (CoA, absea#PQ-843) for lot-specific data including purity assessment (typically by SDS-PAGE), identity confirmation, and endotoxin levels where applicable.

Troubleshooting common issues:

• Protein aggregation: If precipitation is observed, centrifuge the solution at low speed (500–1,000 × g, 5 min, 4°C) and use the supernatant. Aggregation may result from freezing at incorrect concentrations or the absence of carrier protein.
• Loss of activity: Verify storage conditions and confirm that freeze-thaw cycles have been minimized. Ensure reconstitution buffer is appropriate and fresh.
• Low signal in binding assays: Optimize protein coating concentration, incubation temperature, and duration. Verify that detection reagents and assay formats are compatible with the protein tag or native form used.
• Contamination: Discard the vial and contact Aladdin Scientific technical support if microbial contamination is suspected.

For further technical assistance, contact Aladdin Scientific customer support, referencing SKU rp368052 and lot number from the CoA.

Frequently asked questions

What expression system is used to produce Recombinant Human GPR158 Protein (rp368052)?

The specific expression system used for rp368052 should be confirmed by consulting the Certificate of Analysis (CoA, absea#PQ-843) or contacting Aladdin Scientific technical support. Common systems for membrane-associated GPCRs include mammalian cell expression (e.g., HEK293) or insect cell systems (e.g., Sf9), which better preserve post-translational modifications relevant to GPCR function.

Can Recombinant Human GPR158 Protein be used in cell-based functional assays?

Recombinant GPR158 protein in soluble or partially purified form is primarily intended for biochemical assays such as ligand binding, protein interaction studies, and immunoassays. For cell-based functional assays measuring downstream signaling (e.g., cAMP, calcium flux), overexpression of GPR158 in appropriate cell lines is typically preferred. Consult Aladdin Scientific technical support to clarify the form and suitability of rp368052 for your specific assay format.

How should I handle Recombinant Human GPR158 Protein to prevent loss of activity?

To preserve activity, equilibrate the vial to room temperature before opening, reconstitute gently without vortexing, and aliquot immediately into single-use volumes for storage at -80°C. Avoid repeated freeze-thaw cycles, and consider adding 0.1% BSA as a carrier protein when working at low concentrations. Always follow the specific instructions provided in the CoA for rp368052.

Is Recombinant Human GPR158 Protein (rp368052) suitable for use as an ELISA standard?

Recombinant proteins can be used as standards or positive controls in ELISA assays, provided the protein is recognized by the detection antibody used in the assay. Confirm the compatibility of rp368052 with your antibody pair by consulting the CoA and, if needed, contact Aladdin Scientific technical support for guidance on antigen-antibody compatibility.

References

1. Orlandi C, et al. GPR158 mediates RGS7 function and interacts with RGS7-Gβ5 complexes. Research describing GPR158's interaction with RGS7 scaffolding complexes and its role in neuronal signaling.
2. Hauser AS, et al. Pharmacogenomics of GPCR Drug Targets. Cell. 2017. Review covering the landscape of GPCR targets including orphan receptors such as GPR158.
3. Aladdin Scientific Product Page: Recombinant Human GPR158 Protein, SKU rp368052. Certificate of Analysis absea#PQ-843.

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