Recombinant Human GPR179 Protein: Research Guidelines

Recombinant Human GPR179 Protein is a research-grade recombinant protein targeting the orphan G protein-coupled receptor GPR179, a critical component of the retinal ON-bipolar cell signaling cascade implicated in congenital stationary night blindness.

Product Name

Recombinant Human GPR179 Protein

SKU

rp366773

Protein Family

Orphan G protein-coupled receptor (Class A GPCR)

Primary Research Application

Retinal ON-bipolar cell signaling; congenital stationary night blindness (CSNB) research

Species Origin

Human (Homo sapiens)

Recommended Storage

Store at -80°C; avoid repeated freeze-thaw cycles

Quality Standard

Research-grade recombinant protein; purity and activity details per lot certificate of analysis

Background and Biological Significance of GPR179

GPR179 is an orphan G protein-coupled receptor (GPCR) that plays an essential role in retinal signal transduction. It is specifically expressed in ON-bipolar cells of the retina, where it is required for the proper localization and function of the mGluR6 signaling complex at the dendritic tips. Loss-of-function mutations in the GPR179 gene have been identified as a cause of autosomal recessive congenital stationary night blindness (CSNB), a non-progressive retinal disorder characterized by impaired scotopic and photopic vision.

Structurally, GPR179 belongs to the Class A GPCR superfamily and is considered an orphan receptor because no endogenous ligand has been definitively characterized. Its interaction with regulators of G protein signaling (RGS) proteins, particularly RGS7 and RGS11, at the ON-bipolar cell dendrite is critical for the recovery kinetics of the photoresponse. Recombinant Human GPR179 Protein enables researchers to study these interactions, screen for potential ligands, and investigate the molecular mechanisms underlying CSNB.

Protein Specifications and Quality Attributes

Recombinant Human GPR179 Protein (SKU: rp366773) is produced as a recombinant protein derived from the human GPR179 gene sequence. As with all recombinant GPCR-derived proteins, researchers should consult the product's Certificate of Analysis (CoA) for lot-specific information on purity, molecular weight, formulation buffer, endotoxin levels, and biological activity.

Key quality attributes to verify upon receipt include:

• Purity: Assessed by SDS-PAGE and/or HPLC; confirm the expected band or peak corresponding to the target molecular weight.
• Endotoxin level: Critical for cell-based assays; confirm that endotoxin content meets the requirements of your specific application.
• Biological activity: Confirm activity data provided in the CoA are relevant to your planned assay format.
• Formulation: Note the buffer composition and any carrier proteins (e.g., BSA) that may interfere with downstream assays.

For lot-specific data, always request or download the CoA from Aladdin Scientific at the time of purchase.

Reconstitution, Handling, and Storage Guidelines

Proper reconstitution and storage are essential to maintain the activity and integrity of Recombinant Human GPR179 Protein. Follow these general guidelines, and always defer to the specific instructions provided in the product datasheet or CoA:

1. Reconstitution: If supplied in lyophilized form, briefly centrifuge the vial before opening to collect the powder. Reconstitute in the recommended buffer (typically sterile PBS or the manufacturer-specified buffer) to the target concentration. Allow the protein to dissolve gently; avoid vigorous vortexing.
2. Aliquoting: Prepare single-use aliquots immediately after reconstitution to minimize degradation from repeated freeze-thaw cycles. Label each aliquot with date and concentration.
3. Short-term storage: Reconstituted protein may be stored at 4°C for short-term use (typically up to 1 week, depending on formulation). Verify with the CoA.
4. Long-term storage: Store lyophilized or aliquoted reconstituted protein at -80°C. Adding a carrier protein such as 0.1% BSA may enhance stability for some recombinant proteins; confirm suitability with the CoA.
5. Freeze-thaw cycles: Limit to no more than 2-3 cycles. Repeated freeze-thaw significantly reduces protein activity and increases aggregation risk.

Handle all recombinant proteins on ice during experimental setup to minimize proteolytic degradation.

Experimental Applications and Assay Guidance

Recombinant Human GPR179 Protein is suitable for a range of in vitro research applications. Researchers should validate the protein for each specific application and optimize conditions accordingly:

• Protein-protein interaction studies: Use in pull-down assays, co-immunoprecipitation (co-IP), or surface plasmon resonance (SPR) to characterize GPR179 interactions with RGS7, RGS11, or other binding partners in the ON-bipolar cell signaling complex.
• Antibody validation: Use as a positive control antigen in Western blotting, ELISA, or immunofluorescence to validate anti-GPR179 antibodies.
• Structural and biophysical studies: Suitable as a reference standard in assays investigating GPCR folding, stability, or ligand-binding capacity.
• Ligand screening: Employ in binding assays to identify potential endogenous or synthetic ligands for this orphan receptor.
• Functional assays: Incorporate into cell-free or reconstituted signaling systems to study G protein coupling or downstream effector activation.

Always include appropriate positive and negative controls in each experiment. Optimize protein concentration, incubation time, and buffer conditions empirically for each assay format.

Troubleshooting Common Issues

The following troubleshooting guidance addresses frequently encountered issues when working with Recombinant Human GPR179 Protein:

Issue	Likely Cause	Recommended Action
No or weak signal in Western blot	Protein degradation or insufficient loading	Verify storage conditions; increase protein amount; confirm antibody compatibility
Protein aggregation after reconstitution	Incorrect reconstitution buffer or temperature	Use manufacturer-recommended buffer; reconstitute at 4°C; centrifuge at low speed to remove aggregates
High background in ELISA	Endotoxin contamination or non-specific binding	Confirm endotoxin level from CoA; optimize blocking conditions; use additional washes
Loss of activity after storage	Repeated freeze-thaw or improper temperature	Aliquot upon first thaw; store at -80°C; add carrier protein if recommended
Inconsistent results between lots	Lot-to-lot variability	Always validate new lots against previous lot using the same assay; request bridging data from Aladdin Scientific

Contact Aladdin Scientific technical support if issues persist after implementing the above recommendations.

Frequently asked questions

What is GPR179 and why is it important in retinal research?

GPR179 is an orphan G protein-coupled receptor expressed in ON-bipolar cells of the retina. It plays a critical role in organizing the mGluR6 signaling complex and recruiting RGS proteins required for normal photoresponse recovery. Mutations in GPR179 cause autosomal recessive congenital stationary night blindness (CSNB), making it a key target for vision research.

How should Recombinant Human GPR179 Protein (rp366773) be stored to maintain activity?

The protein should be stored at -80°C for long-term preservation. Upon first thaw, prepare single-use aliquots to avoid repeated freeze-thaw cycles, which degrade protein activity. For short-term use, the reconstituted protein may be kept at 4°C for up to one week; always confirm exact conditions with the lot-specific Certificate of Analysis.

What applications is Recombinant Human GPR179 Protein validated for?

Recombinant Human GPR179 Protein is suitable for protein-protein interaction studies, antibody validation (Western blot, ELISA, immunofluorescence), structural and biophysical analyses, orphan receptor ligand screening, and functional signaling assays. Researchers should independently validate the protein for each specific application and optimize experimental conditions accordingly.

How do I reconstitute lyophilized Recombinant Human GPR179 Protein?

Briefly centrifuge the vial before opening to collect the lyophilized powder. Add the manufacturer-recommended reconstitution buffer (typically sterile PBS) to achieve the desired concentration. Mix gently by pipetting or slow rotation; do not vortex vigorously. Allow the protein to fully dissolve on ice before use or aliquoting.

Can Recombinant Human GPR179 Protein be used as an antigen for antibody production?

Recombinant proteins such as GPR179 can in principle be used as antigens for antibody generation; however, the suitability of this specific product for immunization protocols depends on its purity, folding state, and formulation. Consult the product CoA and contact Aladdin Scientific technical support for guidance on antigen preparation for immunization.

References

1. Peachey NS, et al. GPR179 is required for depolarizing bipolar cell function and is mutated in autosomal-recessive complete congenital stationary night blindness. Am J Hum Genet. 2012.
2. Orlandi C, et al. Anchoring of the mGluR6 signaling complex at the ON-bipolar cell dendritic tips requires GPR179. J Neurosci. 2013.
3. Aladdin Scientific. Recombinant Human GPR179 Protein (rp366773) Product Datasheet and Certificate of Analysis. Aladdin Scientific.

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