OR10G4 Antibody

What is OR10G4 and what is its biological function?

OR10G4 (Olfactory receptor 10G4) belongs to the large family of G-protein-coupled receptors (GPCRs) arising from single coding-exon genes. These receptors interact with odorant molecules in the nose to initiate neuronal responses that trigger smell perception. The olfactory receptor gene family is the largest in the genome, with OR10G4 specifically functioning in the recognition and G protein-mediated transduction of odorant signals . The receptor is particularly notable for its role in the perception of compounds such as guaiacol, with genetic variations affecting perception intensity and valence .

What is the molecular structure and cellular localization of OR10G4?

OR10G4 shares the characteristic 7-transmembrane domain structure common to olfactory receptors and many neurotransmitter and hormone receptors. It is primarily localized in the cell membrane as a multi-pass membrane protein . The protein has a molecular weight of approximately 34-35 kDa as observed in experimental detection . This structure facilitates its role in binding odorant molecules and initiating signal transduction cascades leading to olfactory perception.

What are the common identifiers and aliases for OR10G4?

When searching databases or literature, researchers should be aware of several common identifiers for OR10G4:

These identifiers are essential for accurate database searches and consistent reporting in research communications .

What types of OR10G4 antibodies are available for research?

Researchers have access to both monoclonal and polyclonal antibodies against OR10G4. Monoclonal antibodies, such as those derived from mouse (e.g., YP-mAb-13441), offer high specificity and consistency between batches . Polyclonal antibodies, often produced in rabbit (e.g., SAB4501706), typically recognize multiple epitopes on the target protein, potentially providing stronger signals in certain applications . Selection should be based on the specific experimental requirements and the degree of specificity needed.

How should I validate the specificity of an OR10G4 antibody?

Validation of OR10G4 antibody specificity requires a multi-faceted approach:

• Western blot analysis: Confirm the detection of bands at the expected molecular weight (34-35 kDa) in positive control lysates from cells known to express OR10G4 (such as HeLa, COLO, or MCF-7 cells) .

• Blocking peptide competition: Perform parallel experiments where one sample is treated with the synthesized peptide used as the immunogen. Disappearance of signal in the blocked sample confirms specificity .

• Multiple detection methods: Validate across different techniques (Western blot, immunofluorescence, ELISA) to ensure consistent results.

• Negative controls: Include samples from tissues or cell lines that do not express OR10G4 to confirm absence of non-specific binding.

• Genetic validation: When possible, use samples with OR10G4 knockdown or from subjects with known genetic variations to further confirm specificity.

What are the recommended dilutions and conditions for OR10G4 antibodies in common applications?

Based on manufacturer specifications and research protocols, the following dilutions are recommended:

These ranges provide starting points, but researchers should perform dilution series to determine optimal conditions for their specific antibody lot and experimental system.

How do I optimize Western blot conditions for OR10G4 detection?

Optimizing Western blot conditions for OR10G4 requires attention to several key factors:

• Sample preparation: Use fresh lysates from cells with known OR10G4 expression (HeLa, COLO, MCF-7) . Include protease inhibitors to prevent degradation.

• Protein loading: Load 20-50 μg of total protein per lane, adjusting based on expression levels.

• Membrane selection: PVDF membranes may provide better results than nitrocellulose for this hydrophobic transmembrane protein.

• Blocking conditions: Use 5% non-fat dry milk or BSA in TBST. For phospho-specific detection, BSA is preferred.

• Antibody incubation: Start with manufacturer-recommended dilutions in blocking buffer. Incubate primary antibody at 4°C overnight for optimal binding.

• Detection method: For low abundance, consider using high-sensitivity chemiluminescence substrates or fluorescent secondary antibodies.

• Expected result: Look for a specific band at approximately 34-35 kDa .

Why might I observe multiple bands or no signal when detecting OR10G4 by Western blot?

Multiple bands or no signal in OR10G4 Western blots can result from several factors:

Multiple bands may indicate:

• Post-translational modifications (glycosylation, phosphorylation)

• Protein degradation (add fresh protease inhibitors)

• Antibody cross-reactivity with similar olfactory receptors

• Presence of splice variants or polymorphic forms

Absent signal may result from:

• Low expression levels in sample (consider using positive control lysates)

• Inefficient protein transfer

• Suboptimal antibody dilution

• Degraded antibody (check storage conditions)

• Sample preparation issues affecting epitope accessibility

• Genetic variations in your sample affecting antibody binding site

Troubleshooting should begin with positive controls like HeLa, COLO, or MCF-7 cell lysates, which have been documented to express detectable levels of OR10G4 .

How can I assess the quality and functionality of OR10G4 antibodies?

Quality assessment should include:

• Titration experiments: Perform dilution series to determine optimal antibody concentration.

• Reproducibility testing: As demonstrated in avidity assays, coefficient of variation for intraplate reproducibility should ideally be below 10%, with interplate reproducibility between 99-108% .

• Specificity controls: Use peptide blocking, knockout/knockdown samples, or comparative analysis with multiple antibodies targeting different epitopes.

• Lot-to-lot consistency: When changing lots, perform side-by-side comparisons using the same samples and protocols.

• Storage stability assessment: Follow manufacturer recommendations (typically -20°C for 1 year) and avoid repeated freeze-thaw cycles.

How do genetic polymorphisms in OR10G4 affect antibody binding and experimental design?

Genetic polymorphisms in OR10G4 present significant challenges and considerations for antibody-based research:

The human OR10G4 gene exhibits high genetic variability, with multiple common alleles in the population. Research has shown that 63% of studied odorant receptors have polymorphisms that alter in vitro function, and on average, two individuals differ functionally at over 30% of their odorant receptor alleles .

For OR10G4 specifically, variations in genotype have been demonstrated to explain over 15% of observed variation in perceived intensity and over 10% of observed variation in perceived valence for high-affinity agonists like guaiacol .

Implications for experimental design:

• Epitope considerations: Select antibodies targeting conserved regions of OR10G4 to minimize the impact of polymorphisms.

• Validation across genotypes: When possible, validate antibody binding across samples with known OR10G4 genotypes.

• Genotyping subjects: For functional studies correlating antibody binding with perceptual phenotypes, genotype subjects for common OR10G4 variants.

• Control selection: Include controls representing different OR10G4 alleles in comparative studies.

How can I use OR10G4 antibodies to study the relationship between receptor expression and olfactory perception?

Investigating the relationship between OR10G4 expression and perception requires integrating antibody-based techniques with sensory and genetic approaches:

• Tissue expression analysis: Use immunohistochemistry with OR10G4 antibodies to quantify receptor expression in olfactory epithelium biopsies from subjects with different perceptual phenotypes.

• Correlation studies: Combine Western blot quantification of OR10G4 in olfactory tissue with psychophysical testing of specific odorants like guaiacol.

• Genetic-functional integration: Genotype subjects for known OR10G4 polymorphisms (particularly those affecting amino acids 230-279, which contains important functional domains) , then correlate with antibody-detected expression levels and perceptual data.

• Functional imaging: Pair antibody-based receptor quantification with fMRI studies during odor presentation to connect receptor expression with neural activation patterns.

• In vitro models: Develop cell culture systems expressing different OR10G4 variants, using antibodies to confirm expression before functional assays.

Research has demonstrated that variations in OR10G4 genotype explain significant variation in the perception of specific compounds, particularly guaiacol (>15% of intensity variation), but not for lower-affinity agonists like vanillin and ethyl vanillin .

What is known about OR10G4's role in odor perception and how is it being studied?

Recent research has provided significant insights into OR10G4's role in olfactory perception:

Studies have identified specific agonists for OR10G4, including guaiacol, vanillin, and ethyl vanillin. Importantly, research has established that OR10G4 shows higher affinity for guaiacol compared to the other compounds .

Multiple common alleles of OR10G4 exist in the human population, including variants with different functional properties. Some alleles show functional impairment that cannot be attributed to a single SNP, suggesting complex interactions between multiple residues .

Current research methodologies include:

• Heterologous expression systems to assess receptor function in vitro

• Correlation of genotype with perceptual phenotypes

• Antibody-based detection of expression patterns

• Functional studies of receptor-ligand interactions

How can antibodies help elucidate the mechanisms of OR10G4-mediated signal transduction?

OR10G4 antibodies are valuable tools for investigating signal transduction mechanisms:

• Protein-protein interaction studies: Use co-immunoprecipitation with OR10G4 antibodies to identify interacting G proteins and other signaling partners in native tissue or heterologous systems.

• Trafficking and internalization: Apply immunofluorescence with OR10G4 antibodies to track receptor localization changes upon odorant binding and subsequent signaling.

• Post-translational modifications: Employ phospho-specific antibodies alongside general OR10G4 antibodies to monitor signaling-dependent modifications.

• Conformational changes: Develop conformation-specific antibodies that selectively recognize active versus inactive receptor states.

• Quantitative analysis: Use ELISA-based methods with OR10G4 antibodies to quantify receptor levels in different functional states or genetic backgrounds.

These approaches can help resolve how genetic variations in OR10G4, particularly those that affect the response to specific odorants like guaiacol, alter the downstream signaling cascade and ultimately influence perceptual differences between individuals .