OR5M11 Antibody
Description
Introduction to Olfactory Receptor 5M11 Antibody
The OR5M11 Antibody is a specialized research tool developed for detecting and studying the Olfactory receptor 5M11 (OR5M11) protein. This polyclonal antibody is primarily produced in rabbits through immunization with synthetic peptides derived from specific regions of the human OR5M11 protein . The antibody exhibits strong reactivity with human samples and has been validated for multiple laboratory applications .
As an immunological reagent, the OR5M11 Antibody plays a crucial role in advancing our understanding of olfactory receptors and their functions. The antibody enables researchers to visualize, quantify, and characterize OR5M11 expression patterns across various tissues and experimental conditions, providing valuable insights into the molecular mechanisms underlying the sense of smell .
The development and characterization of antibodies targeting specific olfactory receptors like OR5M11 represent significant contributions to sensory biology research, potentially leading to advancements in areas including fragrance development, food science, and disease diagnosis .
Target Protein: Olfactory Receptor 5M11
The target of this antibody, Olfactory Receptor 5M11 (OR5M11), belongs to the G-protein coupled receptor 1 protein family and functions as an olfactory receptor involved in detecting and responding to specific odor molecules . In humans, the canonical OR5M11 protein consists of approximately 305 amino acid residues with a molecular mass of 34.5 kDa .
OR5M11 is primarily localized in the cell membrane where it participates in GPCR signaling pathways essential for the sensory perception of smell . Like other olfactory receptors, OR5M11 is believed to operate by binding specific odor molecules, triggering conformational changes that activate intracellular signaling cascades ultimately resulting in odor perception.
The protein may undergo various post-translational modifications, including glycosylation, which potentially influence its function and localization . Alternative names for this protein include Olfactory receptor 5M11, Olfactory receptor, family 5, subfamily M, member 11, OR11-199, and OR5MB .
Antibody Production and Purification
The OR5M11 Antibody is typically generated by immunizing rabbits with synthetic peptides derived from specific regions of the human OR5M11 protein. Most commercial antibodies are developed using peptides from either the C-terminal region or internal sequences of the human OR5M11 protein . This approach ensures high specificity for the target protein.
Following antibody production, the OR5M11 Antibody undergoes purification through affinity chromatography using epitope-specific immunogens . This purification process significantly enhances the antibody's specificity by removing non-specific antibodies, resulting in a high-quality reagent suitable for sensitive applications.
Physical and Chemical Properties
The OR5M11 Antibody is typically supplied in liquid form with specific buffer components designed to maintain stability and activity. The standard formulation generally includes:
| Component | Purpose | Typical Concentration |
|---|---|---|
| PBS | Buffer | pH 7.4, 150 mM NaCl |
| Glycerol | Stabilizer | 50% |
| BSA | Carrier protein | 0.5% |
| Sodium azide | Preservative | 0.02% |
The antibody concentration typically ranges around 1 mg/ml, though this may vary between suppliers . This formulation ensures antibody stability during shipping, storage, and experimental use.
Western Blotting
Western blotting represents one of the primary applications for the OR5M11 Antibody. This technique allows researchers to detect and semi-quantitatively analyze OR5M11 protein expression in cell or tissue lysates. Recommended dilution ranges for Western blotting applications typically fall between 1:500 and 1:2000, though some suppliers suggest ranges as broad as 1:100 to 1:3000 .
A Western blot analysis using the OR5M11 Antibody can reveal the presence and relative abundance of the target protein while confirming its molecular weight. Documentation from antibodies-online shows successful application in HuvEc cells, demonstrating the antibody's effectiveness in this common technique .
Immunofluorescence and Immunocytochemistry
The OR5M11 Antibody has been validated for immunofluorescence (IF) and immunocytochemistry (ICC) applications, enabling visualization of OR5M11 protein localization within cells and tissues. For these applications, recommended dilution ranges typically fall between 1:200 and 1:1000 .
These techniques provide valuable insights into the subcellular localization of OR5M11, its expression patterns across different cell types, and potential co-localization with other proteins. The high specificity of the antibody makes it suitable for detecting the target protein in fixed cells and tissue sections with minimal background signal.
Enzyme-Linked Immunosorbent Assay
Enzyme-Linked Immunosorbent Assay (ELISA) represents another validated application for the OR5M11 Antibody. This quantitative technique allows precise measurement of OR5M11 protein levels in various samples. For ELISA applications, recommended dilutions are typically higher than for other techniques, with most suppliers suggesting dilutions around 1:5000 .
Handling Precautions
Several important handling precautions should be observed when working with the OR5M11 Antibody:
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Avoid repeated freeze-thaw cycles, which can compromise antibody activity and reduce performance .
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Aliquot the antibody into smaller volumes before freezing to minimize freeze-thaw cycles .
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Handle with care as the product contains sodium azide, which is a poisonous and hazardous substance that should be handled by trained personnel only .
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Follow standard laboratory safety procedures and wear appropriate personal protective equipment when working with the antibody.
Working Solution Preparation
For experimental applications, the OR5M11 Antibody should be diluted to appropriate working concentrations in suitable buffers. The optimal dilution depends on the specific application and should be determined empirically, though manufacturers provide recommended ranges as starting points .
Olfactory System Research
The OR5M11 Antibody serves as a valuable tool for investigating the expression and function of OR5M11 in the olfactory system. By enabling specific detection of this receptor, the antibody contributes to research exploring how OR5M11 participates in odor detection and discrimination .
The olfactory receptor family, to which OR5M11 belongs, plays a critical role in our ability to detect and distinguish thousands of different odors. Research using the OR5M11 Antibody can help identify the specific odor molecules that activate this receptor, potentially enhancing our understanding of the molecular basis of smell perception .
Sensory Biology and Signal Transduction
As a G-protein coupled receptor involved in sensory perception, OR5M11 represents an interesting subject for studying signal transduction mechanisms. The OR5M11 Antibody enables researchers to investigate how this receptor couples to intracellular signaling pathways and how these signals are processed to generate sensory perceptions .
Understanding the signaling properties of specific olfactory receptors like OR5M11 can provide insights into how the brain interprets chemical signals from the environment, translating molecular interactions into conscious perceptions of smell.
Potential Applications in Fragrance and Food Science
Research on olfactory receptors has significant implications for fragrance development and food science. By identifying the specific odor molecules that activate OR5M11 receptors, researchers may contribute to the development of new fragrances and food flavors with specific olfactory properties .
The OR5M11 Antibody enables studies examining how this receptor interacts with various odorant compounds, potentially leading to novel applications in industries where scent and flavor play important roles.
Implications for Health and Disease
Research into olfactory receptors may also have implications for health and disease. The OR5M11 Antibody can support investigations into how alterations in olfactory receptor expression or function contribute to disorders affecting the sense of smell .
Additionally, olfactory dysfunction can serve as an early indicator of certain neurodegenerative diseases. The ability to study specific olfactory receptors like OR5M11 at the molecular level may contribute to developing diagnostic tools and therapeutic approaches for conditions affecting olfactory function.
Product Specs
Target Background
Q&A
What is OR5M11 and why are antibodies against it important for research?
OR5M11 (Olfactory receptor 5M11) is a G-protein coupled receptor primarily involved in olfactory signaling pathways and sensory perception of smell . Though initially characterized as an odorant receptor, emerging research has revealed its unexpected role in pathogen-host interactions, making it a target of growing scientific interest .
OR5M11 is a 305-amino acid protein with a molecular weight of approximately 34.5 kDa that localizes to the cell membrane as a multi-pass membrane protein . The protein undergoes post-translational modifications, including glycosylation, which likely impacts its function and interactions .
Antibodies against OR5M11 are crucial research tools that enable:
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Detection and quantification of endogenous OR5M11 expression across different tissues
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Investigation of OR5M11's role in both olfactory signaling and pathogen interactions
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Functional blocking studies to determine the protein's mechanistic contributions to various cellular processes
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Verification of protein-protein interactions involving OR5M11
What are the key specifications to consider when selecting an OR5M11 antibody?
When selecting an OR5M11 antibody for research applications, several critical specifications should be evaluated to ensure optimal performance:
Research applications should be prioritized when selecting an antibody, as different experimental questions may require targeting specific domains of OR5M11. For instance, studies focused on the protein's role in pathogen interactions might benefit from antibodies targeting the extracellular domains, while investigations of signaling might require antibodies recognizing intracellular regions .
How should OR5M11 antibodies be validated for research applications?
Proper validation of OR5M11 antibodies is crucial for ensuring reliable experimental results. A comprehensive validation approach should include:
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Specificity testing:
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Reproducibility assessment:
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Testing across multiple cell lines or tissue samples
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Evaluation by different researchers or laboratories
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Analysis using different lots of the antibody when possible
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Functional validation:
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Blocking experiments to confirm the antibody's ability to inhibit OR5M11 function
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Research has demonstrated that pretreatment with OR5M11-specific antibodies can significantly inhibit A. pleuropneumoniae adherence to cells expressing OR5M11, confirming both antibody specificity and functional blocking capacity
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Application-specific validation:
A well-documented example of functional validation comes from the study where an OR5M11-specific antibody was used to block the interaction between bacterial Adh protein and OR5M11, resulting in significantly reduced IL-8 secretion (p < 0.05) .
How does OR5M11 antibody blocking affect bacterial pathogen interactions and immune responses?
Research has uncovered a fascinating role for OR5M11 in bacterial pathogenesis, particularly in the context of Actinobacillus pleuropneumoniae infections. OR5M11 antibodies have proven valuable in elucidating these mechanisms:
The interaction between A. pleuropneumoniae adhesin (Adh) and OR5M11 on host cells triggers significant immune responses, particularly the secretion of IL-8 . Specifically:
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OR5M11 expression increased the adherence of A. pleuropneumoniae to 293T cells by approximately 30% (p < 0.05)
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Pretreatment with an OR5M11-specific antibody significantly inhibited:
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A. pleuropneumoniae adherence to Adh-expressing 293T cells (p < 0.05)
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A. pleuropneumoniae wild-type-mediated IL-8 release (p < 0.05)
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Control experiments using isotype control antibodies showed no inhibition of bacterial adherence (p < 0.05)
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The OR5M11-specific antibody had no effect on IL-8 release mediated by A. pleuropneumoniae with Adh deletion (5b ΔAdh)
These findings demonstrate that OR5M11 antibodies can modulate pathogen-host interactions by blocking receptor-adhesin binding, subsequently affecting downstream immune responses. This represents a potential therapeutic approach for bacterial infections where similar interactions occur, and provides a methodological framework for investigating receptor-pathogen interactions in other systems.
What methodological approaches optimize OR5M11 antibody performance in different experimental contexts?
Optimizing OR5M11 antibody performance requires tailored approaches for different experimental applications:
Western Blot Optimization:
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Sample preparation considerations:
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Use fresh tissue/cell lysates with protease inhibitors to prevent OR5M11 degradation
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For membrane proteins like OR5M11, optimize lysis buffers to effectively solubilize the protein (e.g., RIPA buffer with 0.1% SDS)
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Heat samples at lower temperatures (37°C instead of 95°C) to prevent aggregation of transmembrane proteins
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Protocol adjustments:
Immunofluorescence Optimization:
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Fixation methods:
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4% paraformaldehyde fixation preserves membrane structures
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Avoid methanol fixation which can disrupt membrane protein epitopes
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Gentle permeabilization with 0.1% Triton X-100 or 0.1% saponin
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Signal enhancement strategies:
ELISA Applications:
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For quantification assays:
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For interaction studies:
How can OR5M11 antibodies be utilized in studying the dual roles of olfactory receptors in sensory and non-sensory functions?
The discovery that OR5M11, canonically an olfactory receptor, plays a role in pathogen recognition highlights the emerging understanding that olfactory receptors have functions beyond their classical roles in odorant detection. OR5M11 antibodies provide powerful tools for investigating this functional duality:
Comparative Expression Analysis:
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Tissue distribution mapping:
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Use western blot and immunohistochemistry with OR5M11 antibodies to quantify expression across olfactory and non-olfactory tissues
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Correlate expression levels with functional readouts in different cell types
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Compare with other olfactory receptors to identify patterns in non-canonical expression
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Subcellular localization studies:
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Employ immunofluorescence with OR5M11 antibodies to determine if subcellular localization differs between olfactory neurons and immune cells
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Co-localization studies with organelle markers to reveal potential differences in trafficking
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Functional Mechanistic Studies:
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Signaling pathway differentiation:
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Use OR5M11 antibodies in immunoprecipitation experiments to identify different binding partners in olfactory versus immune cells
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Blocking studies to determine if antibody inhibition affects olfactory and immune functions differently
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Compare downstream signaling cascades activated by OR5M11 in different cellular contexts
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Ligand-binding competition assays:
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Develop assays where OR5M11 antibodies compete with odorants and pathogen components for receptor binding
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Determine if different epitope-targeting antibodies differentially affect odorant versus pathogen binding
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Investigate whether post-translational modifications of OR5M11 affect antibody binding and function in different contexts
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This research area has significant implications for understanding the evolutionary repurposing of sensory receptors for immune functions, and OR5M11 antibodies provide crucial tools for dissecting these dual roles with molecular precision.
What is the relationship between OR5M11 and immune responses, and how can antibodies help investigate this connection?
The unexpected finding that OR5M11 interacts with bacterial adhesins and influences immune responses opens an exciting research avenue. OR5M11 antibodies can be instrumental in elucidating this relationship:
Interaction Mapping Studies:
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Determining binding domains:
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Use different antibodies targeting specific epitopes of OR5M11 to block pathogen binding
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Create epitope mapping experiments to identify critical regions for pathogen recognition
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Compare these regions with those involved in odorant binding
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Structural studies:
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Employ antibodies as crystallization chaperones for structural determination of OR5M11
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Use Fab fragments of OR5M11 antibodies to stabilize the protein for cryo-EM studies
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Develop conformation-specific antibodies to capture different activation states
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Immune Response Monitoring:
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Cytokine profiling:
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Monitor changes in IL-8 secretion and other cytokines following OR5M11 antibody treatment
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Research has demonstrated that blocking OR5M11 with specific antibodies significantly reduces IL-8 secretion in response to A. pleuropneumoniae infection (p < 0.05)
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Expand studies to examine broader cytokine profiles and immune cell activation
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Receptor internalization and trafficking:
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Track OR5M11 internalization following pathogen binding using antibody labeling
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Compare trafficking patterns after odorant versus pathogen stimulation
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Investigate whether receptor recycling differs between these contexts
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Translational Applications:
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Diagnostic potential:
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Develop OR5M11 antibody-based assays to detect receptor upregulation during infection
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Correlate OR5M11 expression with disease severity or progression
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Investigate OR5M11 as a biomarker for specific bacterial infections
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Therapeutic implications:
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Explore whether OR5M11 blocking antibodies could serve as adjuvant therapy for specific infections
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Investigate if pathogen binding to OR5M11 could be exploited for targeted drug delivery
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Develop screening assays for compounds that selectively block pathogen-OR5M11 interactions without affecting olfactory function
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These studies could reveal evolutionary connections between olfactory sensing and immune recognition, with OR5M11 potentially representing an ancient mechanism for environmental surveillance that has been repurposed for pathogen detection.
