OR2AJ1 Antibody

What is OR2AJ1 and what is its function in human biology?

OR2AJ1 (Olfactory Receptor Family 2 Subfamily AJ Member 1) is a protein encoded by the OR2AJ1 gene in humans. This receptor belongs to the largest gene family in the genome - the olfactory receptor family. Functionally, OR2AJ1 is a G-protein-coupled receptor (GPCR) with a characteristic 7-transmembrane domain structure that interacts with odorant molecules in the nasal epithelium . When an odorant binds to this receptor, it initiates a neuronal response cascade that ultimately triggers the perception of specific smells. The protein has a molecular weight of approximately 42 kDa and shares structural similarities with other neurotransmitter and hormone receptors .

What experimental applications are OR2AJ1 antibodies validated for?

OR2AJ1 antibodies have been validated for multiple research applications, with specific validation depending on the antibody source. Most commercially available OR2AJ1 antibodies are validated for:

Western blot analyses have demonstrated specific detection of OR2AJ1 in human cell lines including Jurkat, HUVEC, and HT-29 cells . When selecting an antibody, researchers should verify the validation status for their specific application and cell/tissue type of interest .

How should OR2AJ1 antibodies be stored and handled for optimal performance?

For most OR2AJ1 antibodies, the recommended storage conditions are:

• Long-term storage: -20°C for up to one year from the date of receipt

• Short-term storage (after reconstitution): 4°C for up to one month

• Avoid repeated freeze-thaw cycles as this can compromise antibody activity

Most commercially available OR2AJ1 antibodies come in liquid form in PBS containing 50% glycerol, 0.5% BSA, and 0.02% sodium azide at pH 7.4 . Some antibodies may be provided in lyophilized form and require reconstitution with distilled water prior to use . Always refer to the lot-specific information provided by the manufacturer for precise storage and handling instructions.

What epitopes are targeted by commercial OR2AJ1 antibodies?

Commercial OR2AJ1 antibodies are generally raised against synthetic peptides derived from specific regions of the human OR2AJ1 protein. The most common epitope regions include:

These epitope selections are strategically chosen to avoid the transmembrane regions, focusing instead on more accessible regions that maintain native conformation in the folded protein . The immunogen specificity is particularly important when studying closely related olfactory receptors, as cross-reactivity can compromise experimental results.

What cell and tissue types express OR2AJ1 and are suitable for antibody validation?

While olfactory receptors are primarily expressed in olfactory sensory neurons, research has demonstrated that OR2AJ1 can also be detected in non-olfactory tissues and cell lines. Validated cell lines for OR2AJ1 antibody testing include:

• Jurkat cells (human T lymphocyte cells)

• HUVEC cells (human umbilical vein endothelial cells)

• HT-29 cells (human colorectal adenocarcinoma cells)

Western blot analysis shows specific bands at the expected molecular weight of 42 kDa in these cell lines. When implementing OR2AJ1 antibodies in new cell types or tissues, researchers should include appropriate positive controls from these validated sources .

How does OR2AJ1 compare to other olfactory receptors in the same subfamily?

OR2AJ1 belongs to the OR2 subfamily of olfactory receptors, which includes other members such as OR2A1, OR2AG1, and OR2AG2. While they share the characteristic 7-transmembrane GPCR structure, they differ in their amino acid sequences and potentially in their odorant binding profiles.

When conducting comparative studies of these receptors, researchers should be careful to select antibodies with demonstrated specificity to avoid cross-reactivity, particularly when the proteins share high sequence homology in certain regions .

What methodological considerations are important when optimizing Western blot protocols for OR2AJ1 detection?

For optimal Western blot detection of OR2AJ1:

• Sample preparation: Total protein extraction from validated cell lines (Jurkat, HUVEC, HT-29) should include appropriate protease inhibitors to prevent degradation of the target protein.

• Blocking optimization: Commercial validation data shows best results when using 5% non-fat milk or BSA in TBST for blocking, with some antibodies showing cleaner results with BSA.

• Antibody dilution: Start with the manufacturer's recommended dilution range (typically 1:500-1:1000) and optimize based on signal-to-noise ratio. For Real-Gene Labs antibody, a dilution of 1:500-1:1000 is recommended for Western blot applications .

• Specific controls: Include both positive controls (validated cell lines) and negative controls (peptide blocking). Several antibody suppliers demonstrate specificity by showing that the signal disappears when the antibody is pre-incubated with the immunizing peptide .

• Band verification: The expected molecular weight for OR2AJ1 is approximately 42 kDa. Any additional bands should be carefully investigated for potential splice variants or post-translational modifications.

What approaches can resolve cross-reactivity issues between OR2AJ1 and similar olfactory receptors?

Cross-reactivity is a significant concern when studying olfactory receptors due to their high sequence homology. To address this:

• Epitope analysis: Perform sequence alignment analyses between OR2AJ1 and related receptors (particularly OR2A1, OR2AG1, and OR2AG2) to identify regions with lowest homology. Select antibodies targeting these unique regions.

• Validation by knockout/knockdown: Implement siRNA knockdown or CRISPR-Cas9 knockout of OR2AJ1 to confirm antibody specificity. Disappearance of the signal in knockout samples provides strong evidence of specificity.

• Immunoprecipitation followed by mass spectrometry: This approach can conclusively identify the proteins being recognized by the antibody beyond just molecular weight confirmation.

• Peptide competition assays: As demonstrated in the validation data for some commercial antibodies, pre-incubation with the immunizing peptide should abolish specific signals while leaving non-specific signals intact .

• Multi-antibody approach: Utilize multiple antibodies targeting different epitopes of OR2AJ1 to corroborate findings and increase confidence in specificity.

How can OR2AJ1 antibodies be employed in studying the role of this receptor in non-olfactory tissues?

Recent proteogenomic research suggests that olfactory receptors may have functions beyond their canonical roles in olfactory sensory neurons . To investigate OR2AJ1 in non-olfactory contexts:

• Tissue microarray screening: Employ OR2AJ1 antibodies in immunohistochemistry screening across multiple tissue types to identify novel sites of expression.

• Co-immunoprecipitation studies: Use OR2AJ1 antibodies to pull down the receptor and its interacting partners, followed by mass spectrometry to identify tissue-specific signaling complexes.

• Subcellular localization: Combine OR2AJ1 antibodies with organelle markers in confocal microscopy to determine precise subcellular localization, which may provide clues to non-canonical functions.

• Functional assays: Following identification of OR2AJ1 expression in specific non-olfactory cell types, design functional assays (calcium imaging, cAMP production) to test receptor activation by potential ligands.

• Single-cell analysis: Implement OR2AJ1 antibodies in single-cell proteomics workflows to identify rare cell populations expressing this receptor in heterogeneous tissues.

What considerations are important when using OR2AJ1 antibodies in proteogenomic research approaches?

The application of OR2AJ1 antibodies in proteogenomic studies requires specific methodological considerations:

• Antibody specificity validation: Before integration into large-scale proteogenomic workflows, rigorous validation of antibody specificity is essential using methods described in question 3.2.

• Sample preparation optimization: Membrane proteins like OR2AJ1 require specialized extraction protocols to maintain native conformation. Detergent selection is critical, with options including NP-40, Triton X-100, or specialized membrane protein extraction kits.

• Integration with transcriptomic data: As seen in the proteogenomic analysis of antibody-secreting cells , correlating protein-level detection (via antibodies) with transcript-level expression can help identify discrepancies that might indicate post-transcriptional regulation.

• Statistical analysis of cross-platform data: When integrating antibody-based detection with other -omic platforms, statistical methods should account for the different dynamic ranges and technical variabilities of each platform.

• Pathway analysis: Contextualizing OR2AJ1 expression within broader signaling networks requires integration of antibody-based detection with pathway analysis tools to generate testable hypotheses about the receptor's function in specific cellular contexts.