OR10G7 Antibody

What is OR10G7 and why is it significant in research contexts?

OR10G7 (Olfactory Receptor Family 10 Subfamily G Member 7) is a protein encoded by the OR10G7 gene in humans. It belongs to the large family of G-protein-coupled receptors (GPCRs) that typically function as olfactory receptors in the nose . Interestingly, emerging research indicates that olfactory receptors, including OR10G7, are expressed in non-olfactory tissues and may have physiological functions beyond smell perception . This expanding role makes OR10G7 significant for research into cellular signaling pathways, particularly in skin cells and potentially other tissues. The receptor has a molecular weight of approximately 34kDa and features the characteristic seven-transmembrane domain structure common to GPCRs .

What are the common applications for OR10G7 antibodies in research?

OR10G7 antibodies have been validated for several research applications:

Researchers primarily use these antibodies to detect endogenous levels of OR10G7 protein in human samples, study its expression patterns across different tissues, and investigate its potential role in physiological processes like cell proliferation and signaling pathways .

How should researchers optimize Western blot protocols for OR10G7 detection?

For optimal Western blot detection of OR10G7:

• Sample preparation: Use RIPA buffer containing protease inhibitor cocktail for tissue or cell lysis .

• Protein loading: Load 20-40 μg of total protein per lane.

• Electrophoresis: Use 10-12% SDS-PAGE gels for effective separation around the 34 kDa range .

• Transfer: PVDF membranes are recommended for optimal protein binding .

• Blocking: Use 5% non-fat milk or BSA in TBST for 1 hour at room temperature.

• Primary antibody incubation: Dilute OR10G7 antibody 1:500-1:2000 in blocking buffer and incubate overnight at 4°C .

• Detection: Anti-rabbit IgG HRP-linked secondary antibodies are suitable for visualization .

Note that multiple bands may be observed in Western blots for OR10G7, potentially due to post-translational modifications including glycosylation, acetylation, and methylation .

How can researchers validate the specificity of OR10G7 antibodies?

Validating OR10G7 antibody specificity requires multiple complementary approaches:

• Positive control selection: HUVEC cells have demonstrated endogenous OR10G7 expression and can serve as positive controls .

• Peptide competition assay: Pre-incubate the antibody with the immunizing peptide (amino acids 224-273 of human OR10G7) to confirm signal specificity .

• siRNA knockdown verification: Compare signal between control and OR10G7 siRNA-treated samples to confirm specificity .

• Multiple antibody validation: Use antibodies from different sources or raised against different epitopes of OR10G7.

• Recombinant expression systems: Overexpression of OR10G7 in cell lines can provide additional confirmation of antibody specificity.

In which human tissues has OR10G7 expression been confirmed?

Research suggests that OR10G7, while originally characterized in olfactory neurons, shows expression in various non-olfactory tissues:

• Confirmed expression: Human keratinocytes, particularly HaCaT cell lines

• Potential expression: Other skin cells, based on studies of related olfactory receptors (ORs)

• Variable expression: Different studies suggest that expression patterns of ORs, including OR10G7, may be tissue-specific and potentially context-dependent

For comprehensive tissue expression analysis, researchers should employ multiple detection methods including RT-qPCR, Western blotting, and immunohistochemistry/immunofluorescence to verify expression patterns across different cell types .

How does OR10G7 function relate to cell signaling pathways?

Based on research on related olfactory receptors, OR10G7 likely functions through canonical G-protein signaling pathways:

• G-protein coupling: As a GPCR, OR10G7 likely couples to G proteins upon activation .

• Calcium signaling: Studies on related ORs suggest involvement in modulating intracellular calcium levels, which can affect multiple downstream pathways .

• MAPK pathway activation: Related ORs have been shown to activate ERK1/2 signaling .

• Proliferation effects: Related ORs like OR7A17 influence cell proliferation pathways, suggesting OR10G7 may have similar functions .

Research on OR7A17, another olfactory receptor, shows that it influences calcium influx and promotes keratinocyte proliferation, potentially providing a model for understanding OR10G7 function .

How might OR10G7 be involved in dermatological research and skin biology?

Emerging evidence suggests OR10G7 and related olfactory receptors may play significant roles in skin biology:

• Cell proliferation regulation: Related receptor OR7A17 has been implicated in keratinocyte proliferation, suggesting OR10G7 might have similar functions .

• Retinoic acid response: Studies on OR7A17 show downregulation by all-trans retinoic acid (ATRA), affecting cell proliferation through calcium signaling pathways .

• Potential therapeutic target: The involvement of ORs in proliferation suggests they could be targets for treating proliferative skin disorders .

• Expression in atopic dermatitis: Related OR10G7 has been implicated in atopic dermatitis, suggesting potential roles in inflammatory skin conditions .

Researchers investigating OR10G7 in skin biology should consider its potential interactions with retinoic acid signaling pathways and effects on cell proliferation, similar to what has been observed with OR7A17 .

What experimental approaches are recommended for studying OR10G7 ligands and activation?

Identifying ligands and studying OR10G7 activation requires specialized techniques:

• Calcium imaging assays: Monitor intracellular calcium fluctuations in OR10G7-expressing cells when exposed to potential ligands .

• cAMP accumulation assays: Measure changes in cAMP levels as indicators of OR10G7 activation.

• Reporter gene assays: Use luciferase or GFP reporters downstream of OR10G7 activation to screen potential ligands.

• Overexpression systems: Generate stable cell lines overexpressing OR10G7 to facilitate activation studies .

• Knock-down/Knock-out approaches: Use siRNA or CRISPR-Cas9 to study loss-of-function effects .

• Structural modeling: Employ computational approaches to predict ligand binding sites based on the seven-transmembrane domain structure common to GPCRs .

For comprehensive characterization, researchers should combine multiple approaches to validate ligand identification and activation mechanisms.

How can researchers address multiple band detection in Western blots for OR10G7?

Multiple bands in OR10G7 Western blots are common and may represent:

• Post-translational modifications: OR10G7 may undergo glycosylation, acetylation, or methylation, resulting in multiple bands of different molecular weights .

• Validation approach: Researchers should verify which band represents OR10G7 through:

  • siRNA knockdown to identify which bands decrease

  • Overexpression systems to identify which bands increase

  • Peptide competition assays to determine which bands are blocked by the immunizing peptide

• Sample preparation: Modify lysis conditions to preserve or disrupt specific modifications

• Gradient gels: Use 4-20% gradient gels for better resolution of closely spaced bands

As noted in one study: "the appearance of several bands for OR10G7 might be attributed to post-translational modifications such as glycosylation, acetylation, and methylation" .

What are recommended storage and handling procedures to maintain OR10G7 antibody activity?

For optimal results with OR10G7 antibodies:

• Storage conditions:

  • Store at -20°C upon receipt

  • Avoid repeated freeze/thaw cycles

  • Consider aliquoting the antibody upon first thaw

• Working solution preparation:

  • Dilute in appropriate buffer with 0.5% BSA immediately before use

  • Most OR10G7 antibodies are supplied in PBS containing 50% glycerol, 0.02% sodium azide, and may contain 0.5% BSA

• Shipping considerations:

  • Antibodies are typically shipped at 4°C

  • Transfer to -20°C storage immediately upon receipt

• Stability:

  • Follow manufacturer's expiration dates

  • Monitor performance if using beyond recommended shelf life

Following these guidelines will help ensure consistent experimental results and extend the useful life of the antibody.

How does OR10G7 research compare with studies on other olfactory receptors in non-olfactory tissues?

Several olfactory receptors have been studied in non-olfactory contexts, providing comparative insights:

Research on OR7A17 is particularly relevant as it shows that olfactory receptor expression in keratinocytes can influence proliferation and respond to retinoic acid signaling , potentially providing a model for understanding OR10G7 function.

What research methodologies are emerging for studying GPCR-related signaling of OR10G7?

Advanced methodologies for investigating OR10G7 signaling include:

• BRET/FRET assays: These fluorescence-based techniques can monitor real-time protein-protein interactions involved in OR10G7 signaling.

• Phospho-specific antibodies: To detect activation of downstream signaling components such as ERK1/2, JNK1/2/3, and PLC-γ1 .

• Single-cell calcium imaging: For high-resolution analysis of OR10G7-mediated calcium responses.

• Receptor trafficking analysis: Using fluorescently tagged OR10G7 to monitor internalization and recycling.

• Transcriptomics approaches: RNA-seq analysis to identify genes regulated downstream of OR10G7 activation.

• Proteomics: Mass spectrometry-based approaches to identify interaction partners and post-translational modifications.

• CRISPR-Cas9 gene editing: For generating knockout or knockin models to study OR10G7 function .