OR10C1 Antibody

What is OR10C1 and why is it studied in research?

OR10C1 (Olfactory receptor family 10 subfamily C member 1) is a G-protein-coupled receptor (GPCR) that belongs to the large family of olfactory receptors. These receptors interact with odorant molecules in the nose to initiate neuronal responses that trigger the perception of smell. OR10C1 has a characteristic 7-transmembrane domain structure common to many neurotransmitter and hormone receptors. The olfactory receptor gene family is the largest in the genome, making these proteins important subjects for studying GPCR functionality, signal transduction, and sensory perception mechanisms . The protein has several alternative names including hs6M1-17, OR10C1P, and OR10C2 .

Which applications are most commonly validated for OR10C1 antibodies?

Based on validated antibody data, OR10C1 antibodies have been successfully employed in multiple experimental applications, primarily:

The most consistently validated application across providers is Western Blot, suggesting this may be the most reliable technique for OR10C1 detection .

How do I select the appropriate OR10C1 antibody for my experiment?

When selecting an OR10C1 antibody, consider these key criteria:

• Experimental application: Choose antibodies specifically validated for your intended application (WB, ICC, IHC, or ELISA). For instance, if performing Western blot analysis, consider antibodies like Invitrogen PA5-102114 or GeneTex GTX87088 which are specifically validated for this technique .

• Antibody type: Polyclonal antibodies like Affinity Biosciences DF5006 provide higher sensitivity through multiple epitope recognition, while monoclonal antibodies offer higher specificity for single epitopes.

• Species reactivity: Verify that the antibody recognizes OR10C1 in your experimental species. Most validated OR10C1 antibodies are optimized for human samples .

• Validation data: Prioritize antibodies with published validation data showing specificity for OR10C1. Leading validated antibodies include those from Affinity Biosciences, antibodies-online, LSBio, Invitrogen, and GeneTex .

• Epitope information: When available, select antibodies targeting specific regions of interest on the OR10C1 protein. For instance, some antibodies like the Invitrogen PA5-102114 target a specific peptide derived from human OR10C1 corresponding to amino acid residues H243-N293 .

What are the optimal protocols for Western blot detection of OR10C1?

When performing Western blot for OR10C1 detection, follow these methodological guidelines:

• Sample preparation:

  • For tissue samples: Homogenize in RIPA buffer with protease inhibitors

  • For cell samples: Lyse directly in sample buffer with reducing agent

• Gel percentage recommendation:

  • Use 10-12% SDS-PAGE gels as OR10C1 is approximately 35-40 kDa

• Antibody dilutions:

  • Primary OR10C1 antibody: 1:1000 dilution is typically effective for validated antibodies like the Invitrogen PA5-102114

  • Secondary antibody: 1:5000 anti-rabbit IgG-HRP for rabbit polyclonal primaries

• Blocking conditions:

  • 5% non-fat dry milk in TBST (Tris-buffered saline with 0.1% Tween-20) for 1 hour at room temperature

  • Alternative: 3% BSA in TBST when using phospho-specific antibodies

• Detection strategy:

  • Enhanced chemiluminescence (ECL) for standard detection

  • Longer exposure times (1-5 minutes) may be necessary for detecting lower expression levels

• Expected molecular weight:

  • Confirm bands at approximately 35-40 kDa for OR10C1

  • Be aware of potential post-translational modifications that may alter migration

How can I validate OR10C1 antibody specificity in my experimental system?

Comprehensive validation of OR10C1 antibody specificity requires multiple complementary approaches:

• Positive and negative control samples:

  • Positive controls: Tissues known to express OR10C1 (olfactory epithelium)

  • Negative controls: Tissues with minimal OR10C1 expression or OR10C1 knockout models

• Peptide competition assay:

  • Pre-incubate the OR10C1 antibody with excess synthesized peptide derived from the human OR10C1 protein (such as the H243-N293 peptide region used for Invitrogen antibody generation)

  • Run parallel assays with blocked and unblocked antibody

  • Specific binding should be significantly reduced in the peptide-blocked sample

• Recombinant protein expression:

  • Overexpress tagged OR10C1 protein in a cell line with low endogenous expression

  • Confirm antibody detection of both endogenous and overexpressed protein

  • Verify molecular weight shifts with fusion tags

• siRNA/shRNA knockdown:

  • Transiently reduce OR10C1 expression in expressing cell lines

  • Western blot or immunostaining signal should decrease proportionally

• Multi-antibody concordance:

  • Compare staining patterns from multiple OR10C1 antibodies targeting different epitopes (e.g., compare results from Affinity Biosciences DF5006 and GeneTex GTX87088)

  • Consistent results across antibodies suggest higher specificity

• Mass spectrometry validation:

  • Immunoprecipitate OR10C1 using the antibody

  • Confirm protein identity via mass spectrometry

What are the challenges in detecting OR10C1 in tissue samples and how can they be overcome?

Detecting OR10C1 in tissue samples presents several challenges with specific solutions:

• Low expression levels:

  • Challenge: OR10C1, like many olfactory receptors, has restricted expression patterns

  • Solution: Use signal amplification techniques such as tyramide signal amplification (TSA) to enhance detection sensitivity

  • Solution: Consider RNAscope to correlate protein detection with mRNA expression

• Specificity concerns:

  • Challenge: Cross-reactivity with other olfactory receptor family members due to sequence homology

  • Solution: Perform appropriate controls using tissues from OR10C1 knockout models

  • Solution: Use antibodies validated against peptide sequences unique to OR10C1

• Tissue preparation issues:

  • Challenge: Epitope masking during fixation, particularly problematic for membrane proteins

  • Solution: Test multiple fixation protocols (4% PFA, methanol, acetone) to identify optimal conditions

  • Solution: Include antigen retrieval steps (citrate buffer pH 6.0 or EDTA buffer pH 8.0)

• Background staining:

  • Challenge: Non-specific binding in olfactory tissues

  • Solution: Increase blocking time (overnight at 4°C) with 3-5% normal serum

  • Solution: Add 0.1-0.3% Triton X-100 for better antibody penetration

• Antibody optimization:

  • Challenge: Determining optimal antibody concentration

  • Solution: Perform titration experiments with dilution series (1:100 to 1:2000)

  • Solution: Extend primary antibody incubation to 48-72 hours at 4°C for tissues

• Signal verification:

  • Challenge: Confirming true positive signal

  • Solution: Use fluorescent co-localization with olfactory neuron markers

  • Solution: Compare with in situ hybridization data for OR10C1 mRNA

How can I design co-localization studies using OR10C1 antibodies with other cellular markers?

Effective co-localization studies with OR10C1 antibodies require careful experimental design:

• Selection of compatible antibodies:

  • Choose OR10C1 antibodies raised in different host species than your other markers

  • For example, pair rabbit polyclonal OR10C1 antibodies like Invitrogen PA5-102114 with mouse monoclonal antibodies against other targets

  • Verify that secondary antibodies don't cross-react

• Marker selection strategy:

  • Membrane markers: Pair with other GPCR antibodies or membrane markers to confirm surface localization

  • Cellular compartment markers: Use antibodies against ER (calnexin), Golgi (GM130), or early endosomes (EEA1) to track processing and trafficking

  • Cell-type markers: Combine with olfactory neuron markers like OMP (Olfactory Marker Protein) or Golf (olfactory G protein)

• Sequential immunostaining protocol:

  • Block with 5% normal serum + 0.3% Triton X-100 for 2 hours

  • Incubate with first primary antibody overnight at 4°C

  • Wash extensively (5 × 10 minutes)

  • Apply first secondary antibody (2 hours, room temperature)

  • Wash extensively (5 × 10 minutes)

  • Block again for 1 hour

  • Apply second primary antibody overnight at 4°C

  • Wash and apply second secondary antibody

  • Include appropriate controls for each antibody separately

• Advanced imaging considerations:

  • Use confocal microscopy with sequential scanning to prevent bleed-through

  • Apply deconvolution algorithms to improve signal resolution

  • Implement Pearson's correlation coefficient or Mander's overlap coefficient to quantify co-localization

• Technical controls:

  • Single antibody controls to establish baseline signals

  • Secondary-only controls to identify non-specific binding

  • Signal intensity normalization between channels

What analytical approaches can detect potential OR10C1 post-translational modifications?

Investigating post-translational modifications (PTMs) of OR10C1 requires specialized approaches:

• Glycosylation analysis:

  • Run parallel Western blots treating samples with PNGase F to remove N-linked glycans

  • Compare migration patterns before and after treatment

  • Use lectins alongside OR10C1 antibodies to confirm glycosylation sites

• Phosphorylation detection:

  • Enrich phosphorylated proteins using phosphoprotein enrichment columns

  • Treat lysates with phosphatase inhibitors during extraction

  • Consider generating phospho-specific OR10C1 antibodies for key regulatory sites

  • Use Phos-tag™ SDS-PAGE to separate phosphorylated from non-phosphorylated forms

• Ubiquitination assessment:

  • Immunoprecipitate OR10C1 using validated antibodies

  • Probe with anti-ubiquitin antibodies on Western blots

  • Include proteasome inhibitors (MG132) in treatment conditions to stabilize ubiquitinated forms

• Palmitoylation investigation:

  • Employ click chemistry approaches with alkyne-modified palmitic acid

  • Use hydroxylamine treatment to cleave thioester bonds of palmitoylated proteins

  • Compare migration patterns before and after treatment

• Mass spectrometry-based PTM mapping:

  • Immunoprecipitate OR10C1 using validated antibodies

  • Digest with multiple proteases to ensure complete sequence coverage

  • Analyze by LC-MS/MS with neutral loss scanning for phosphorylation

  • Search MS data against databases of known PTM patterns

• Mutational analysis validation:

  • Generate site-directed mutants of predicted PTM sites

  • Express in heterologous systems alongside wild-type OR10C1

  • Compare antibody detection, localization, and function

How should I interpret discrepancies between different antibody-based detection methods for OR10C1?

When facing discrepancies between different detection methods, consider these analytical approaches:

What are the most common false positives when detecting OR10C1 and how can they be identified?

False positives in OR10C1 detection can arise from multiple sources:

• Cross-reactivity with other olfactory receptors:

  • Issue: The human genome contains approximately 400 functional olfactory receptor genes with sequence similarity

  • Identification: Compare observed molecular weight with predicted OR10C1 size

  • Verification: Use peptide competition assays with OR10C1-specific peptides

  • Control: Include samples from tissues not expressing OR10C1

• Non-specific binding in Western blots:

  • Issue: Bands appearing at unexpected molecular weights

  • Identification: Compare with positive control sample band patterns

  • Verification: Increase blocking stringency (5% milk to 5% BSA)

  • Control: Include secondary-only controls to identify non-antibody specific bands

• Background in immunostaining:

  • Issue: Diffuse signal not correlating with expected cellular distribution

  • Identification: Compare with negative control tissues and secondary-only controls

  • Verification: Increase washing steps (5 × 10 minutes) with 0.1% Tween-20

  • Control: Pre-absorb secondary antibodies with tissue powder

• Autofluorescence interference:

  • Issue: Endogenous fluorescence misinterpreted as antibody signal

  • Identification: Examine unstained samples for natural fluorescence

  • Verification: Image using multiple channels to identify spectrum of autofluorescence

  • Control: Use Sudan Black B (0.1%) treatment to quench autofluorescence

• Overinterpretation of low-level signals:

  • Issue: Weak signals amplified during image processing

  • Identification: Compare signal-to-noise ratios across samples

  • Verification: Quantify signal intensity relative to background

  • Control: Establish detection thresholds based on negative controls

How can I properly quantify OR10C1 expression levels using antibody-based techniques?

Accurate quantification of OR10C1 requires rigorous methodology:

• Western blot quantification protocol:

  • Load concentration gradient of recombinant OR10C1 to establish standard curve

  • Include housekeeping protein controls (β-actin, GAPDH) for normalization

  • Capture images within linear dynamic range of detection system

  • Use densitometry software with background subtraction

  • Report results as relative expression normalized to controls

• Immunofluorescence quantification approach:

  • Capture images with identical exposure settings across all samples

  • Define regions of interest (ROIs) consistently across samples

  • Measure mean fluorescence intensity minus background

  • Normalize to cell number or area

  • Analyze ≥100 cells per condition for statistical validity

• ELISA-based absolute quantification:

  • Develop sandwich ELISA using two antibodies targeting different OR10C1 epitopes

  • Generate standard curve using recombinant OR10C1 protein

  • Process samples in technical triplicates

  • Include spike-in controls to assess matrix effects

  • Report concentrations in ng/ml or pmol/L

• Flow cytometry quantification:

  • Use antibodies validated for flow cytometry applications

  • Establish negative population gates using isotype controls

  • Report data as percent positive and mean fluorescence intensity

  • Use standardized beads to convert arbitrary units to molecules of equivalent soluble fluorochrome (MESF)

• Statistical analysis requirements:

  • Perform minimum of three biological replicates

  • Apply appropriate statistical tests based on data distribution

  • Report both effect size and p-values

  • Conduct power analysis to ensure adequate sample size

How can OR10C1 antibodies be utilized in receptor trafficking and internalization studies?

Investigating OR10C1 trafficking dynamics requires specialized methodologies:

• Surface biotinylation assay protocol:

  • Culture cells expressing OR10C1

  • Label surface proteins with cell-impermeable NHS-SS-biotin

  • Allow internalization for various time points

  • Strip remaining surface biotin with membrane-impermeable reducing agent

  • Lyse cells and pull down internalized biotinylated proteins

  • Detect OR10C1 with validated antibodies like Invitrogen PA5-102114

  • Quantify internalization rate relative to total surface labeling

• Antibody feeding assay optimization:

  • Generate OR10C1 construct with extracellular epitope tag (FLAG, HA)

  • Apply anti-tag antibody to live cells at 4°C to label surface receptors

  • Warm cells to 37°C to permit internalization

  • Detect internalized antibody-receptor complexes after acid wash

  • Co-stain with endosomal markers to track intracellular trafficking

• Live-cell imaging approach:

  • Create OR10C1-fluorescent protein fusion constructs

  • Validate fusion protein function and localization

  • Perform time-lapse confocal microscopy

  • Calculate diffusion coefficients and mobile fraction

  • Correlate with fixed-cell antibody staining for validation

• Endocytic pathway co-localization:

  • Use pulsed agonist stimulation to trigger receptor internalization

  • Fix cells at various time points post-stimulation

  • Co-stain with markers for early endosomes (EEA1), recycling endosomes (Rab11), and lysosomes (LAMP1)

  • Quantify co-localization coefficients at each time point

  • Create kinetic profile of receptor trafficking through compartments

• Recycling rate quantification:

  • Apply cleavable biotin to surface proteins

  • Allow internalization, then cleave remaining surface biotin

  • Allow recycling at 37°C for various time points

  • Re-apply membrane-impermeable biotin-cleaving reagent

  • Quantify protected (internal) versus cleaved (recycled) fractions

What approaches can resolve controversies regarding OR10C1 tissue expression patterns?

Resolving conflicting reports about OR10C1 expression requires multifaceted approaches:

• Multi-antibody consensus mapping:

  • Compare staining patterns across multiple validated OR10C1 antibodies

  • Test each antibody under identical conditions across tissue panel

  • Prioritize signals detected by multiple independent antibodies

  • Document discrepancies systematically

• Correlative expression analysis:

  • Perform parallel analysis of:

    • Protein detection (immunohistochemistry)

    • mRNA detection (in situ hybridization)

    • Transcript quantification (qPCR)

  • Map concordance and discrepancies between detection methods

  • Weight evidence based on method sensitivity and specificity

• Single-cell resolution techniques:

  • Apply single-cell RNA sequencing to identify cell populations expressing OR10C1

  • Use flow cytometry with validated OR10C1 antibodies to isolate expressing cells

  • Perform dual RNA/protein detection via RNAscope combined with immunofluorescence

  • Correlate expression with cell-type specific markers

• Genetic validation approaches:

  • Generate reporter knockin models (OR10C1 promoter driving fluorescent protein)

  • Use CRISPR-Cas9 to tag endogenous OR10C1 locus

  • Compare reporter expression with antibody detection patterns

  • Document developmental and stimulus-dependent expression changes

• Technical standardization:

  • Develop standard operating procedures for tissue collection and processing

  • Implement positive and negative control tissues in every experiment

  • Use calibrated image acquisition settings across laboratories

  • Establish quantitative thresholds for calling positive expression

• Meta-analysis framework:

  • Systematically compile expression data from published literature

  • Weight studies by methodology robustness

  • Identify patterns in discrepancies (fixation-sensitive epitopes, primer specificity issues)

  • Generate consensus maps with confidence scores for each tissue/cell type

How can I develop a multiplexed assay incorporating OR10C1 antibodies for high-throughput screening?

Developing multiplexed assays with OR10C1 antibodies requires systematic optimization:

• Antibody panel compatibility assessment:

  • Test for cross-reactivity between antibodies in the panel

  • Verify epitope accessibility in multiplexed conditions

  • Optimize antibody concentrations individually and in combination

  • Select antibodies raised in different host species when possible

• Platform selection considerations:

  • Microarray-based: Spot OR10C1 antibodies alongside other targets

  • Bead-based: Conjugate antibodies to distinctly coded microspheres

  • Imaging-based: Use spectrally distinct fluorophores for each antibody

  • Mass cytometry: Label antibodies with distinct metal isotopes for CyTOF analysis

• High-content imaging protocol:

  • Culture cells in 96- or 384-well format

  • Apply test compounds or genetic perturbations

  • Stain with OR10C1 antibody alongside markers for:

    • Cell viability (nuclear dyes)

    • Cellular compartments (organelle markers)

    • Activation states (phospho-specific antibodies)

  • Acquire images via automated microscopy

  • Apply machine learning algorithms for phenotypic classification

• Bead-based multiplex assay development:

  • Conjugate OR10C1 capture antibody to coded microspheres

  • Develop sandwich assay with biotinylated detection antibody

  • Include calibration beads for absolute quantification

  • Validate against single-plex ELISA results

  • Assess cross-reactivity within multiplex environment

• Validation requirements:

  • Determine assay dynamic range, sensitivity, and precision

  • Assess impact of multiplexing on individual antibody performance

  • Evaluate matrix effects using spike-recovery experiments

  • Establish inter- and intra-assay coefficients of variation

  • Compare results with orthogonal single-target methods

What are the optimal strategies for using OR10C1 antibodies in cross-species comparative studies?

Cross-species OR10C1 studies require careful consideration of evolutionary conservation:

• Epitope conservation analysis:

  • Align OR10C1 protein sequences across target species

  • Identify conserved and divergent epitope regions

  • Prioritize antibodies targeting conserved regions for cross-species studies

  • Consider the specific epitope targeted by antibodies like Invitrogen PA5-102114 (H243-N293 region) and assess its conservation

• Validation hierarchy for cross-species applications:

  • Verify antibody reactivity against recombinant OR10C1 from each species

  • Conduct Western blots on tissue lysates from each species

  • Perform peptide competition assays using species-specific peptides

  • Compare staining patterns with mRNA expression data for each species

• Species-specific protocol modifications:

  • Adjust tissue fixation based on species-specific requirements:

    • Rodent tissues: 4% PFA, 4-24 hours

    • Primate tissues: 10% neutral buffered formalin, 24-48 hours

  • Optimize antigen retrieval for each species

  • Adjust antibody concentrations based on signal intensity

• Comparative expression mapping approach:

  • Create standardized tissue panels across species

  • Process and stain tissues in parallel

  • Quantify expression levels using identical imaging parameters

  • Normalize to housekeeping proteins conserved across species

  • Generate comparative expression heat maps

• Functional correlation strategies:

  • Pair antibody detection with functional assays (calcium imaging, cAMP measurements)

  • Compare OR10C1 localization with odorant response profiles

  • Correlate expression patterns with behavioral or physiological responses

  • Document species-specific post-translational modifications

How might OR10C1 antibodies contribute to understanding olfactory receptor trafficking mechanisms?

OR10C1 antibodies offer valuable tools for investigating fundamental questions in olfactory receptor biology:

• Receptor life cycle tracking:

  • Map OR10C1 biosynthesis through secretory pathway using pulse-chase experiments

  • Determine half-life and degradation pathways using validated antibodies

  • Identify quality control checkpoints using co-localization with ER/Golgi markers

  • Correlate trafficking efficiency with functional responses

• Molecular chaperone identification:

  • Perform immunoprecipitation with OR10C1 antibodies

  • Identify interacting proteins via mass spectrometry

  • Validate interactions through reverse co-immunoprecipitation

  • Determine functional significance through chaperone knockdown/overexpression

• Cilia targeting mechanisms:

  • Compare ciliary versus intracellular pools of OR10C1

  • Identify ciliary targeting sequences through mutational analysis

  • Track real-time movement using antibody-based pulse-chase approaches

  • Correlate trafficking efficiency with odorant sensitivity

• Activity-dependent regulation:

  • Monitor OR10C1 localization before and after odorant exposure

  • Quantify surface expression changes following repeated stimulation

  • Distinguish recycling from de novo synthesis pathways

  • Correlate trafficking changes with sensitization/adaptation responses

• Interplay with other olfactory signaling components:

  • Investigate co-trafficking with G proteins and effector enzymes

  • Determine if OR10C1 forms oligomeric complexes

  • Assess impact of accessory proteins on trafficking efficiency

  • Compare trafficking mechanisms across different olfactory receptor subtypes

What novel approaches could overcome current limitations in OR10C1 antibody specificity?

Advancing OR10C1 antibody technology requires innovative approaches:

• Epitope-focused antibody development:

  • Generate antibodies against unique OR10C1 regions with minimal homology to other ORs

  • Create peptide libraries spanning unique regions of OR10C1

  • Screen against OR10C1 and closely related receptors to identify discriminating epitopes

  • Implement negative selection strategies during antibody development

• Conformation-specific antibody generation:

  • Develop antibodies recognizing active versus inactive OR10C1 conformations

  • Stabilize specific receptor conformations using nanobodies or conformation-selective ligands

  • Screen for antibodies that preferentially bind specific functional states

  • Validate using mutagenesis of conformational switches

• Recombinant antibody engineering:

  • Create phage-display libraries for selecting high-affinity OR10C1-specific binders

  • Engineer single-chain variable fragments (scFvs) for improved tissue penetration

  • Generate bispecific antibodies targeting OR10C1 and cell-type markers

  • Design intrabodies for tracking intracellular OR10C1 populations

• Native-conformation immunization strategies:

  • Immunize with cells expressing OR10C1 to preserve native membrane orientation

  • Use proteoliposomes containing purified OR10C1

  • Develop DNA immunization protocols expressing OR10C1 in vivo

  • Select antibodies through cell-based screening to ensure native recognition

• Cross-validation frameworks:

  • Implement systematic cross-validation between antibody-based and genetic approaches

  • Develop standard reference materials for antibody validation

  • Establish open-access databases of validated staining patterns

  • Create community standards for OR10C1 antibody validation reporting

How can OR10C1 antibodies facilitate investigation of ectopic olfactory receptor expression?

OR10C1 antibodies enable exploration of non-canonical expression and function:

• Systematic tissue screening approach:

  • Create tissue microarrays from multiple organ systems

  • Apply validated OR10C1 antibodies like those from the top validated list

  • Correlate protein detection with transcriptomic data

  • Distinguish between basal expression and disease-associated upregulation

• Single-cell resolution methodology:

  • Combine OR10C1 antibody staining with single-cell RNA sequencing

  • Identify rare cell populations expressing OR10C1

  • Correlate expression with cell lineage markers

  • Track expression changes during development and disease progression

• Functional investigation protocol:

  • Isolate OR10C1-expressing cells from non-olfactory tissues using antibody-based sorting

  • Characterize signaling pathways using phospho-specific antibodies

  • Perform calcium imaging to assess ligand responses

  • Compare functional properties between canonical and ectopic expression sites

• Disease-association studies:

  • Screen tissue samples from various pathological conditions

  • Quantify OR10C1 expression changes relative to healthy controls

  • Correlate expression with disease progression markers

  • Investigate potential as diagnostic or prognostic biomarker

• Developmental regulation mapping:

  • Track OR10C1 expression across embryonic and postnatal development

  • Identify developmental windows of expression in non-olfactory tissues

  • Correlate with tissue differentiation markers

  • Investigate potential roles in tissue patterning and development