OR10AG1 Antibody

What is OR10AG1 and what types of antibodies target this protein?

OR10AG1 (Olfactory Receptor, Family 10, Subfamily AG, Member 1) is a human olfactory receptor protein with a molecular weight of approximately 34 kDa. It is also known by alternative names including Olfactory receptor OR11-160 . OR10AG1 antibodies are immunological reagents raised against specific epitopes of this protein, typically produced in rabbits and available in various formats:

• Host species: Predominantly rabbit-derived polyclonal antibodies

• Clonality: Both polyclonal (most common) and monoclonal options are available

• Immunogen regions: Several epitope regions are targeted, including amino acids 214-263, 231-280, and 235-263

• Conjugations: Available as unconjugated or conjugated to detection molecules such as PE (phycoerythrin), biotin, or APC (allophycocyanin)

The diverse range of available antibodies reflects the varying experimental needs in OR10AG1 research, from basic protein detection to complex immunological assays.

How should researchers design experiments with OR10AG1 antibodies to ensure valid results?

Experimental design when working with OR10AG1 antibodies requires careful planning to ensure reliable, reproducible results. Consider implementing these methodological approaches:

• Establish a clear hypothesis: Begin with a specific, testable hypothesis about OR10AG1 that guides your experimental approach .

• Use a randomized block design: When comparing OR10AG1 expression across different experimental conditions, implement a randomized block design to control for confounding variables .

• Include proper controls: Essential controls include:

  • Positive control (tissue/cells known to express OR10AG1)

  • Negative control (tissue/cells known not to express OR10AG1)

  • Isotype controls (rabbit IgG at matching concentrations)

  • Peptide competition assays to verify antibody specificity

• Consider between-subjects vs. within-subjects design: For studies examining OR10AG1 under multiple conditions, carefully choose between comparing different samples (between-subjects) or the same samples under different treatments (within-subjects) .

• Determine sample size: Ensure adequate statistical power through appropriate sample size calculations based on expected effect sizes .

How do epitope differences affect OR10AG1 antibody performance and selection?

The choice between OR10AG1 antibodies targeting different epitopes (amino acid regions) can significantly impact experimental outcomes:

When selecting an OR10AG1 antibody, researchers should consider:

• Protein structure: C-terminal antibodies (like those targeting AA 235-263) may perform better if N-terminal regions are obscured in the native protein conformation.

• Post-translational modifications: Modifications near certain epitopes may affect antibody binding. Review the target region for potential phosphorylation sites or other modifications.

• Splice variants: Different epitope-targeting antibodies may detect different isoforms of OR10AG1.

• Experimental application: Some epitope regions may perform better in certain applications (WB vs. IF) due to differences in protein denaturation and epitope accessibility.

This strategic selection of epitope-specific antibodies can significantly improve detection sensitivity and specificity in OR10AG1 research.

What is the recommended protocol for using OR10AG1 antibodies in Western blotting?

For optimal Western blot results with OR10AG1 antibodies, follow this detailed protocol:

• Sample preparation:

  • Lyse cells or tissues in an appropriate lysis buffer containing protease inhibitors

  • Determine protein concentration using a compatible assay (BCA or Bradford)

  • Prepare samples in 1× Laemmli buffer with reducing agent

  • Heat samples at 95°C for 5 minutes

• Gel electrophoresis:

  • Load 20-50 μg of protein per lane

  • Use a 10-12% polyacrylamide gel (optimal for the 34 kDa OR10AG1 protein)

  • Run at 100-120V until sufficient separation is achieved

• Transfer:

  • Transfer proteins to a PVDF or nitrocellulose membrane

  • Use a wet transfer system at 100V for 1 hour or 30V overnight at 4°C

• Blocking:

  • Block the membrane with 5% non-fat dry milk or BSA in TBST for 1 hour at room temperature

• Primary antibody incubation:

  • Dilute OR10AG1 antibody in blocking buffer at 1:500-1:1000

  • Incubate overnight at 4°C with gentle agitation

• Washing:

  • Wash membrane 3× with TBST, 5-10 minutes each

• Secondary antibody incubation:

  • Use an appropriate anti-rabbit IgG secondary antibody (HRP, AP, or fluorescently labeled)

  • Dilute according to manufacturer's recommendation

  • Incubate for 1 hour at room temperature

• Detection:

  • For HRP-conjugated secondaries, use an enhanced chemiluminescent substrate

  • For fluorescent secondaries, scan with an appropriate imaging system

  • Expected band size for OR10AG1: 34 kDa

This protocol has been validated for detecting OR10AG1 in RAW264.7 cell lysates and can be adapted for other sample types.

What are the optimal storage and handling conditions for maintaining OR10AG1 antibody activity?

Proper storage and handling are crucial for preserving OR10AG1 antibody function and extending shelf life:

• Shipping conditions:

  • OR10AG1 antibodies are typically shipped at 4°C with cool packs

• Long-term storage:

  • Store at -20°C in small aliquots

  • Avoid repeated freeze-thaw cycles as they can cause protein denaturation and loss of activity

• Working solution handling:

  • When preparing diluted working solutions, use sterile tubes and buffers

  • Store diluted antibody at 4°C for short-term use (1-2 weeks)

  • For dilutions containing carrier proteins like BSA, add sodium azide (0.02%) as a preservative

• Formulation considerations:

  • Most OR10AG1 antibodies are supplied in buffers containing glycerol (50%) as a cryoprotectant

  • The standard formulation includes phosphate-buffered saline (PBS) without Mg²⁺ and Ca²⁺, pH 7.4, with 150mM NaCl

• Stability indicators:

  • Monitor for visible precipitation or color changes

  • If protein aggregation is observed, the antibody may have reduced activity

Following these storage guidelines will help ensure consistent performance across experiments and maximize the usable lifetime of OR10AG1 antibodies.

What controls should be included when using OR10AG1 antibodies in immunological assays?

Implementing proper controls is essential for validating results with OR10AG1 antibodies:

• Primary antibody controls:

  • Isotype controls: Use matched rabbit IgG at the same concentration as the OR10AG1 antibody. Products A82272 and A17360 are suitable isotype controls for rabbit polyclonal antibodies .

  • Peptide competition: Pre-incubate the antibody with the immunizing peptide prior to application to confirm specificity. Blocking peptides are available specifically for OR10AG1 antibodies .

• Secondary antibody controls:

  • No primary control: Apply only secondary antibody to identify potential non-specific binding

  • Recommended secondaries: For rabbit-derived OR10AG1 antibodies, suitable secondaries include:

    • Goat Anti-Rabbit IgG H&L Antibody (AP) (A294874)

    • Goat Anti-Rabbit IgG H&L Antibody (Biotin) (A294795)

    • Goat Anti-Rabbit IgG H&L Antibody (FITC) (A294887)

    • Goat Anti-Rabbit IgG H&L Antibody (HRP) (A294888)

• Sample-related controls:

  • Positive tissue/cell control: Include samples known to express OR10AG1

  • Negative tissue/cell control: Include samples known not to express OR10AG1

  • Loading control: Use antibodies against housekeeping proteins (β-actin, GAPDH) to verify equal loading in Western blots

• Technical controls:

  • Serial dilution: Test a range of antibody dilutions to identify optimal signal-to-noise ratio

  • Cross-reactivity assessment: Test the antibody on non-target tissues to assess potential cross-reactivity

Implementing these controls helps distinguish specific from non-specific signals and validates the reliability of OR10AG1 detection in your experimental system.

How can researchers optimize sensitivity when detecting low-abundance OR10AG1 protein?

When working with low-abundance OR10AG1 expression, several strategies can enhance detection sensitivity:

• Sample enrichment techniques:

  • Perform immunoprecipitation to concentrate OR10AG1 before Western blotting

  • Use tissue or cell fractionation to isolate membrane proteins where OR10AG1 is likely to be localized

  • Consider using detergent-resistant membrane preparations to concentrate olfactory receptors

• Signal amplification methods:

  • Employ tyramide signal amplification (TSA) for immunohistochemistry/immunofluorescence

  • Use biotin-streptavidin systems with multiple binding sites for signal enhancement

  • Consider more sensitive detection substrates (e.g., femto-level ECL for Western blots)

• Optimized antibody conditions:

  • Extend primary antibody incubation time (overnight at 4°C or longer)

  • Adjust blocking conditions to reduce background while preserving specific signals

  • Use signal enhancers compatible with your detection system

• Detection system selection:

  • For Western blotting: choose high-sensitivity chemiluminescent substrates

  • For immunofluorescence: select bright, photostable fluorophores and use confocal microscopy

  • For ELISA: consider using amplification steps in the detection protocol

• Validation with multiple antibodies:

  • Use OR10AG1 antibodies targeting different epitopes to confirm results

  • Compare results from antibodies with different conjugations (e.g., unconjugated vs. PE-conjugated)

These strategies can significantly improve the detection of low-abundance OR10AG1 protein while maintaining specificity.

How can OR10AG1 antibodies be integrated into advanced research methodologies?

OR10AG1 antibodies can be leveraged in cutting-edge research approaches:

• High-throughput screening applications:

  • Adapt OR10AG1 antibodies for microarray-based proteomics

  • Develop multiplex assays combining OR10AG1 with other olfactory receptor antibodies

  • Implement automated immunohistochemistry workflows for large-scale tissue analysis

• Advanced imaging techniques:

  • Utilize OR10AG1 antibodies in super-resolution microscopy to examine receptor clustering

  • Apply OR10AG1 antibodies in multiplexed immunofluorescence to study co-localization with interacting proteins

  • Explore live-cell imaging using membrane-permeable fluorescent OR10AG1 antibody fragments

• Single-cell analysis:

  • Integrate OR10AG1 antibodies into single-cell proteomics workflows

  • Develop CyTOF (mass cytometry) applications using metal-conjugated OR10AG1 antibodies

  • Combine with single-cell transcriptomics for multi-omic analysis of olfactory receptor expression

• Conformational studies:

  • Design conformation-specific OR10AG1 antibodies to distinguish active vs. inactive receptor states

  • Implement proximity ligation assays to investigate OR10AG1 protein-protein interactions

  • Use antibodies in structural biology approaches to stabilize OR10AG1 for crystallography

• Therapeutic research applications:

  • Explore OR10AG1 antibodies in receptor internalization and trafficking studies

  • Investigate potential diagnostic applications in olfactory dysfunction research

  • Develop therapeutic antibody derivatives targeting OR10AG1 in relevant pathologies

These advanced applications represent promising future directions for OR10AG1 antibody utilization in scientific research.