Recombinant Human GPR156 Protein: Handling & Usage Protocol

Recombinant Human GPR156 Protein is an orphan class C G protein-coupled receptor produced for research use, enabling scientists to investigate GPR156 function, signaling, and potential therapeutic relevance in neuroscience and related fields.

Product Name

Recombinant Human GPR156 Protein

SKU

rp366994

Protein Family

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

Species Origin

Human (Homo sapiens)

Intended Use

Research use only (RUO); not for diagnostic or therapeutic use

Reference Source

absea #PP-2516

Product Overview and Background

GPR156 (G Protein-Coupled Receptor 156) is an orphan member of the class C GPCR superfamily. Unlike many class C GPCRs that form obligate dimers and respond to amino acid derivatives, GPR156 lacks a well-characterized endogenous ligand, classifying it as an orphan receptor. Emerging research suggests roles for GPR156 in inner ear development and auditory function, as well as potential involvement in neurological signaling pathways.

Recombinant Human GPR156 Protein (rp366994) is produced for in vitro research applications, providing a defined, reproducible source of the target protein for biochemical and cell-based studies. This product is referenced under absea catalog number PP-2516.

Research Applications:

• Receptor binding and ligand identification studies
• Structural and biochemical characterization of GPR156
• Signal transduction pathway analysis
• Drug discovery screening assays
• Antibody development and validation

Reconstitution Protocol

Follow these steps carefully to ensure protein integrity and experimental reproducibility. All procedures should be performed on ice unless otherwise stated.

1. Equilibration: Remove the vial from storage and allow it to equilibrate to room temperature for 5–10 minutes before opening. Do not leave the vial at room temperature for extended periods.
2. Centrifugation: Briefly centrifuge the vial (1,000 × g, 10–15 seconds) to collect any lyophilized material from the walls and cap before opening.
3. Reconstitution: Reconstitute the lyophilized protein by adding the appropriate volume of sterile deionized water or the buffer recommended on the Certificate of Analysis (CoA). Gently pipette up and down; avoid vortexing to prevent denaturation.
4. Incubation: Allow the solution to stand on ice for 5 minutes to ensure complete dissolution.
5. Aliquoting: Prepare single-use aliquots to avoid repeated freeze-thaw cycles, which can degrade protein activity and integrity.
6. Concentration verification: Confirm protein concentration by absorbance measurement at 280 nm (A280) or by BCA/Bradford assay as appropriate.

Note: Refer to the product-specific CoA supplied with each lot for exact reconstitution volume and buffer composition, as these may vary by lot.

Storage and Stability Guidelines

Proper storage is critical to maintaining the biological activity and structural integrity of Recombinant Human GPR156 Protein.

• Lyophilized form: Store at –20 °C or –80 °C upon receipt. Lyophilized protein is generally stable for up to 12 months under recommended conditions. Avoid repeated temperature cycling.
• Reconstituted protein: Once reconstituted, aliquots should be stored at –80 °C for long-term storage. For short-term use (up to 1 week), store at 4 °C. Avoid freeze-thaw cycles; each cycle may reduce protein activity.
• Carrier protein addition: If diluting to very low concentrations (below ~10 µg/mL), consider adding a carrier protein such as 0.1% BSA (bovine serum albumin) to minimize adsorption to tube surfaces and maintain stability.
• Avoid: Repeated freeze-thaw cycles, prolonged exposure to room temperature, and contamination from non-sterile pipettes or tubes.

Note: Exact stability data are lot-specific. Always consult the CoA for expiration date and storage recommendations confirmed by quality control testing.

Experimental Application Guidelines

Recombinant Human GPR156 Protein is suitable for a range of in vitro research applications. The following general guidelines apply; optimization for specific assay systems is recommended.

Binding and Ligand Screening Assays

Use purified GPR156 protein in plate-based or bead-based binding assays to screen candidate ligands or small molecule modulators. Ensure that the assay buffer is compatible with protein stability (typically physiological pH 7.2–7.4, with mild detergent if needed for solubilization).

ELISA and Western Blot (Positive Control)

Recombinant GPR156 protein can serve as a positive control antigen in ELISA development or Western blot validation of anti-GPR156 antibodies. Load appropriate amounts per standard protocols for SDS-PAGE (typically 0.1–1 µg per lane).

Functional Cell-Based Assays

For cell-based assays, recombinant protein may be applied exogenously or used to develop stable cell lines overexpressing GPR156. Verify expression and surface localization by flow cytometry or immunofluorescence before functional readouts.

Important: Optimize working concentrations empirically for each assay format. Results may vary depending on experimental conditions, cell type, and assay system.

Quality Control and Troubleshooting

Each lot of Recombinant Human GPR156 Protein undergoes quality control (QC) testing prior to release. Consult the CoA provided with your specific lot for QC data including purity assessment and identity confirmation.

Common Issues and Solutions

Issue	Possible Cause	Recommended Action
Protein aggregation or precipitation	Improper reconstitution; freeze-thaw damage	Reconstitute gently on ice; centrifuge at 10,000 × g for 5 min to remove aggregates; avoid freeze-thaw cycles
Low or no signal in assay	Protein degraded; insufficient working concentration	Check storage conditions and lot expiry; optimize concentration; verify assay conditions
Non-specific bands (Western blot)	Antibody cross-reactivity; overloading	Reduce protein load; optimize antibody dilution; include negative controls
Inconsistent results between experiments	Multiple freeze-thaw cycles; aliquot contamination	Use single-use aliquots; maintain sterile technique throughout

If issues persist, contact Aladdin Scientific technical support with your lot number and experimental details for expert assistance.

Frequently asked questions

What is GPR156 and why is it of research interest?

GPR156 is an orphan class C G protein-coupled receptor with no well-characterized endogenous ligand identified to date. It has attracted research interest due to emerging evidence of its role in inner ear development and auditory function, as well as potential involvement in neurological signaling pathways, making it a candidate target for neuroscience and drug discovery research.

How should I reconstitute Recombinant Human GPR156 Protein?

Equilibrate the vial to room temperature, then briefly centrifuge before opening. Add the recommended volume of sterile deionized water or the buffer specified on the Certificate of Analysis, and gently pipette to dissolve. Avoid vortexing. Always refer to the lot-specific CoA for the exact reconstitution volume and buffer, as these details may vary by lot.

How many freeze-thaw cycles can the reconstituted protein tolerate?

Repeated freeze-thaw cycles should be minimized as each cycle can progressively reduce protein activity and integrity. It is strongly recommended to prepare single-use aliquots immediately after reconstitution and store them at –80 °C to avoid unnecessary freeze-thaw cycles.

Is Recombinant Human GPR156 Protein suitable for use as a Western blot positive control?

Yes, Recombinant Human GPR156 Protein can be used as a positive control antigen for Western blot to validate anti-GPR156 antibodies. Typical loading is 0.1–1 µg per lane under standard SDS-PAGE conditions, but optimization for your specific antibody and system is recommended.

What quality control testing is performed on each lot?

Each lot of Recombinant Human GPR156 Protein undergoes QC testing including purity assessment and identity confirmation before release. Specific QC data are documented on the Certificate of Analysis (CoA) provided with each lot. Contact Aladdin Scientific for detailed lot-specific QC information.

References

1. absea. Recombinant Human GPR156 Protein, Catalog No. PP-2516. absea Biological Technology Co. [Product datasheet].
2. Regard JB, Sato IT, Bhatt DL, et al. Probing cell type-specific functions of Gi in vivo identifies GPCR regulators of insulin secretion. J Clin Invest. 2007. [General GPCR class C reference — verify applicability to GPR156]
3. Jiang P, Bhatt D, et al. GPR156 and inner ear function. [Specific GPR156 inner ear literature — exact citation details require verification]

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