OR56A3 Antibody

What is OR56A3 and what biological role does it play?

OR56A3 (Olfactory Receptor Family 56 Subfamily A Member 3) is an odorant receptor belonging to the G-protein coupled receptor 1 family. It functions as a multi-pass membrane protein located in the cell membrane. Olfactory receptors interact with odorant molecules in the nose to initiate neuronal responses that trigger smell perception. These receptors share a 7-transmembrane domain structure with many neurotransmitter and hormone receptors and are responsible for the recognition and G protein-mediated transduction of odorant signals . The olfactory receptor gene family is notably the largest in the human genome.

The protein has several alternative names in the literature:

• Olfactory receptor 56A3

• Olfactory receptor 56A6

• Olfactory receptor family 56 subfamily A member 3 pseudogene

• Olfactory receptor OR11-51

• OR56A2P, OR56A3, OR56A3P, OR56A6

What types of OR56A3 antibodies are available for research applications?

Multiple types of OR56A3 antibodies are available for research, with the following specifications:

These antibodies are typically generated against synthetic peptides derived from specific regions of human OR56A3 protein, making them suitable for various experimental applications .

What are the optimal protocols for using OR56A3 antibodies in different applications?

Based on manufacturer recommendations and experimental protocols, the following application guidelines can be followed:

For Immunofluorescence/Immunocytochemistry (IF/ICC):

• Grow cells on coverslips in appropriate media

• Fix cells in 4% paraformaldehyde at 4°C for 20 minutes

• Wash and permeabilize cells with PBS containing 0.1% Triton X-100

• Block in PBS with 1% fish gelatin (or similar blocking agent) for 1 hour at room temperature

• Apply primary OR56A3 antibody at dilution 1:200 - 1:1000 for 2 hours at room temperature

• Wash three times with PBS

• Apply appropriate fluorophore-conjugated secondary antibody for 45 minutes

• Counterstain nuclei with DAPI if desired

• Mount and image using confocal microscopy

For ELISA:

• Use at dilution of 1:20000

• Follow standard ELISA protocols with appropriate positive and negative controls

• Particularly suitable for peptide ELISA formats

For Western Blotting:

• Optimal dilutions should be determined empirically by the end user

• Typically requires optimization between 1:500-1:2000 for most polyclonal antibodies

How should OR56A3 antibodies be stored and handled to maintain efficacy?

For maximum antibody stability and longevity, follow these storage and handling recommendations:

Critical handling considerations:

• Avoid repeated freeze-thaw cycles as they significantly reduce antibody activity

• Store in small aliquots with glycerol (typically provided in 50% glycerol buffer)

• Most preparations contain 0.02% sodium azide as preservative - handle accordingly with appropriate safety precautions

How should researchers design experiments to validate the specificity of OR56A3 antibodies?

A robust validation approach should include multiple complementary methods:

• Recombinant expression system validation:

  • Transfect suitable cell lines (e.g., Hana3A cells) with OR56A3 expression vectors

  • Include tagged version of OR56A3 (e.g., with rhodopsin tag) for co-localization studies

  • Perform immunocytochemistry with both the OR56A3 antibody and antibody against the tag

  • Compare staining patterns between transfected and non-transfected cells

• Control samples testing:

  • Include tissues or cells known to be negative for OR56A3 expression

  • Use blocking peptides (the immunogen peptide) to confirm binding specificity

  • Test antibody on multiple cell types to confirm expected expression patterns

• Multiple antibody approach:

  • When possible, use antibodies targeting different epitopes of OR56A3

  • Compare staining patterns between antibodies from different manufacturers

  • Confirm results using orthogonal methods (e.g., mRNA detection)

What experimental controls are essential when working with OR56A3 antibodies?

These controls help distinguish between specific signals and experimental artifacts, which is critical for accurate data interpretation .

How can OR56A3 antibodies be utilized in cancer research?

Recent findings suggest OR56A3 may have potential implications in cancer biology:

• Expression profiling in tumors:

  • OR56A3 shows no expression in normal breast tissues but variable expression in breast carcinoma tissues and cell lines

  • This differential expression pattern makes it a potential biomarker for specific cancer subtypes

• Comparative expression analysis:

  • Several olfactory receptors (ORs) have been identified as tumor markers

  • OR51E2 and OR51E1 show increased expression in prostate cancer

  • OR51E1 has been postulated as a marker for lung cell carcinoids and cancer of the small intestine

  • OR7C1 is reported as a marker for colon cancer-initiating cells

• Potential research applications:

  • Investigating OR56A3 expression changes during cancer progression

  • Evaluating correlation between OR56A3 expression and clinical outcomes

  • Exploring functional roles of OR56A3 in tumor biology through activation/inhibition studies

  • Developing diagnostic assays based on OR expression profiles

What methodological challenges might researchers encounter when using OR56A3 antibodies?

Common challenges and troubleshooting approaches include:

• Cross-reactivity issues:

  • Olfactory receptors comprise a large gene family with sequence similarities

  • Validate antibody specificity against closely related family members

  • Use epitope mapping to identify unique regions for antibody targeting

• Low signal-to-noise ratio:

  • Optimize blocking conditions (BSA concentration, type of blocking protein)

  • Adjust antibody concentration through careful titration experiments

  • Increase washing stringency to reduce non-specific binding

  • Try alternative detection systems with higher sensitivity

• Inconsistent results between experiments:

  • Standardize protein extraction and sample preparation protocols

  • Use freshly prepared buffers and reagents

  • Document lot numbers of antibodies and evaluate lot-to-lot variability

  • Maintain consistent incubation times and temperatures

• Limited tissue penetration:

  • Optimize fixation protocols (duration, fixative composition)

  • Adjust permeabilization conditions for membrane proteins

  • Consider antigen retrieval methods for formalin-fixed tissues

How do different experimental designs affect OR56A3 antibody performance?

The experimental design significantly impacts antibody performance and data interpretation:

• Sample preparation effects:

  • Fresh vs. frozen vs. fixed samples yield different epitope accessibility

  • Denatured proteins (for WB) may expose different epitopes than native proteins

  • Fixation can mask epitopes or create artificial cross-reactivity

• Detection system considerations:

  • Chromogenic vs. fluorescent detection systems offer different sensitivity ranges

  • Signal amplification methods (e.g., tyramide signal amplification) may be necessary for low abundance targets

  • Multiplex detection requires careful antibody selection to avoid cross-reactivity

• Quantification approach:

  • Western blot densitometry vs. immunofluorescence intensity measurements have different dynamic ranges

  • Flow cytometry offers single-cell resolution but may require different antibody optimization

  • Consider whether relative or absolute quantification is required for the research question

What are new applications of OR56A3 antibodies beyond traditional techniques?

Emerging research methodologies utilizing OR56A3 antibodies include:

• Single-cell analysis:

  • Using OR56A3 antibodies for single-cell protein profiling

  • Combining with single-cell RNA sequencing for multi-omics approaches

  • Spatial transcriptomics with protein validation using OR56A3 antibodies

• High-throughput screening:

  • Development of automated immunoassays for large-scale tissue microarrays

  • Multiplexed antibody panels including OR56A3 for comprehensive OR family profiling

  • Drug screening assays targeting OR56A3 and related receptors

• In vivo imaging:

  • Developing conjugated OR56A3 antibodies for in vivo molecular imaging

  • Using antibody fragments (Fab, scFv) for improved tissue penetration

  • Theranostic applications combining imaging and therapeutic functions

How can researchers address data discrepancies when working with OR56A3 antibodies?

When faced with conflicting data from different OR56A3 antibodies or experimental approaches:

• Comprehensive validation strategy:

  • Use multiple antibodies targeting different epitopes

  • Validate with orthogonal methods (mRNA detection, mass spectrometry)

  • Consider genetic approaches (CRISPR-Cas9) to confirm specificity

• Detailed methodology reporting:

  • Document complete antibody information (manufacturer, catalog number, lot number)

  • Report all experimental conditions in detail (concentration, incubation time, temperature)

  • Share complete protocols to enable reproducibility

• Transparent data analysis:

  • Use appropriate statistical approaches for quantification

  • Report all replicates and variations between experiments

  • Consider interlaboratory validation for critical findings

This systematic approach to addressing discrepancies increases confidence in research findings and advances the field's understanding of OR56A3 biology and function.