IL17RC Antibody

What is IL17RC and what is its role in immune signaling?

IL17RC is a single-pass type I membrane protein that forms part of the receptor complex for the proinflammatory cytokines IL-17A and IL-17F. In humans, the canonical protein has a length of 791 amino acid residues and a mass of 86.2 kDa with subcellular localization in the cell membrane . Unlike IL-17RA, which is predominantly expressed in hemopoietic cells, IL17RC is expressed in nonhemopoietic tissues, particularly prostate, skeletal muscle, kidney, and placenta .

IL17RC functions by forming a heteromeric complex with IL-17RA to mediate signaling from IL-17A and IL-17F homodimers as well as IL-17A/F heterodimers. This signaling is critical for antimicrobial host defense and maintenance of tissue integrity . The cytoplasmic portion contains a SEFIR (SEF/IL-17R) domain that is necessary but not sufficient for signaling - an additional ~20-30 amino acid region downstream (termed "SEFEX" for SEFIR extension) is also required for full functionality .

Functionally, IL17RC is essential for IL-17-dependent immune responses, particularly against fungal infections. Inherited deficiency of IL17RC causes chronic mucocutaneous candidiasis (CMC), characterized by recurrent or persistent infections with Candida species .

What are the common applications for IL17RC antibodies in research?

IL17RC antibodies are utilized across multiple experimental applications:

• Western Blotting (WB): Detects IL17RC protein in cell or tissue lysates, enabling analysis of expression levels, post-translational modifications, and processing. This application helps distinguish between different IL17RC isoforms based on molecular weight .

• Enzyme-Linked Immunosorbent Assay (ELISA): Quantifies IL17RC levels in biological samples or validates antibody specificity against recombinant IL17RC proteins .

• Immunohistochemistry (IHC): Visualizes IL17RC expression patterns in tissue sections, helping determine cellular and subcellular localization. This is particularly valuable for comparing expression across normal and pathological tissues .

• Immunocytochemistry (ICC): Examines IL17RC expression and distribution in cultured cells, allowing for detailed subcellular localization studies .

• Flow Cytometry: Analyzes IL17RC surface expression on individual cells within heterogeneous populations, particularly useful for studying receptor expression dynamics .

• Immunoprecipitation (IP): Isolates IL17RC and its binding partners for further analysis, particularly useful for studying the IL17RA/RC complex and associated signaling molecules like Act1 .

• Neutralization Assays: Blocks IL17RC function to assess the biological consequences of receptor inhibition in experimental systems, especially valuable for functional studies .

What species reactivity is available for IL17RC antibodies?

IL17RC antibodies are available with reactivity against several species, though human and mouse are the most common:

• Human-reactive antibodies:

  • Most commercial antibodies target human IL17RC

  • Human IL17RC has a canonical form of 791 amino acids (86.2 kDa)

  • These antibodies enable studies in human cells, tissues, and disease models

• Mouse-reactive antibodies:

  • Mouse IL17RC antibodies enable research in murine models

  • Mouse IL17RC shares approximately 68-70% amino acid identity with human IL17RC in the extracellular domain

  • Critical for in vivo studies, including infection and autoimmunity models

• Cross-reactive antibodies:

  • Some antibodies recognize conserved epitopes across species

  • Typical cross-reactivity between human and mouse IL17RC is approximately 20%

  • May show varying degrees of cross-reactivity with other mammals

• Ortholog considerations:

  • IL17RC gene orthologs have been reported in mouse, rat, bovine, frog, chimpanzee, and chicken species

  • The degree of conservation affects antibody cross-reactivity

  • Epitope sequence alignment is crucial when using antibodies across species

Researchers should carefully review product data sheets for validated species reactivity and consider testing the antibody on their specific samples from different species when cross-reactivity is critical for comparative studies.

How to validate the specificity of an IL17RC antibody?

Validating IL17RC antibody specificity is crucial for reliable research results and should involve multiple complementary approaches:

• Positive and negative control samples:

  • Test the antibody on cells/tissues known to express IL17RC (positive controls like prostate, skeletal muscle, kidney, placenta)

  • Include IL17RC-negative samples or IL17RC-knockout cells as negative controls

• Antigen competition assays:

  • Pre-incubate the antibody with recombinant IL17RC protein before application

  • Signal reduction or elimination confirms specificity

• RNA interference:

  • Use siRNA or shRNA to knockdown IL17RC expression

  • Reduced antibody signal should correlate with knockdown efficiency

• Multiple antibody validation:

  • Compare results using antibodies targeting different IL17RC epitopes

  • Consistent results increase confidence in specificity

• Correlation with mRNA expression:

  • Compare protein detection with RT-PCR or RNA-seq data

  • Expression patterns should generally correlate

• Glycosylation considerations:

  • IL17RC undergoes glycosylation, which may affect antibody recognition

  • Test detection before and after deglycosylation treatment (e.g., tunicamycin)

• Isoform awareness:

  • With up to 8 different isoforms reported for this protein, understand which isoforms your antibody recognizes

  • Validate against recombinant proteins representing major isoforms

The Human Protein Atlas provides extensive characterization data for each Prestige Antibody target, including IHC tissue arrays of 44 normal human tissues and 20 common cancer types, offering valuable reference data for validation .

What are the key considerations when selecting an IL17RC antibody for flow cytometry versus Western blot?

Selecting the optimal IL17RC antibody requires different considerations depending on the application:

For Flow Cytometry:

• Epitope accessibility:

  • Choose antibodies targeting extracellular epitopes

  • Avoid antibodies against intracellular domains unless using permeabilization

• Native conformation recognition:

  • Select antibodies that recognize native, non-denatured IL17RC

  • Antibodies raised against recombinant full-length protein or extracellular domain fragments are preferable

• Fluorophore selection:

  • Consider directly conjugated antibodies (e.g., Alexa Fluor 488)

  • Select fluorophores compatible with your cytometer and other markers in your panel

• Titration importance:

  • Determine optimal concentration to maximize signal-to-noise ratio

  • Test multiple concentrations (typically 0.1-10 μg/ml)

For Western Blot:

• Denatured epitope recognition:

  • Select antibodies that recognize linear, denatured epitopes

  • Antibodies raised against synthetic peptides often work well

• Isoform detection capabilities:

  • Consider which IL17RC isoforms need to be detected

  • With multiple splice variants, epitope location determines which isoforms will be detected

• Molecular weight awareness:

  • Human canonical IL17RC is 86.2 kDa, but runs at different sizes due to glycosylation

  • Different isoforms have distinct molecular weights

• Post-translational modification sensitivity:

  • Some antibodies may be sensitive to glycosylation states

  • Consider deglycosylation treatments if needed

Common Considerations for Both:

• Validation status:

  • Review literature citations for the specific application

  • Check product data sheets for validation data in your application

• Clonality trade-offs:

  • Monoclonal: Higher specificity, lower sensitivity, single epitope

  • Polyclonal: Higher sensitivity, potential cross-reactivity, multiple epitopes

How do IL17RC antibodies help study the IL-17A/F signaling complex?

IL17RC antibodies provide crucial tools for investigating the IL-17A/F signaling complex through multiple methodological approaches:

• Receptor complex composition analysis:

  • Co-immunoprecipitation with IL17RC antibodies can pull down associated proteins like IL17RA

  • Western blotting of precipitated complexes reveals interaction partners

  • This approach has revealed that IL-17A and IL-17F enhance formation of a multimeric receptor complex containing a specific glycosylated isoform of IL-17RA paired with IL17RC

• Stoichiometry determination:

  • Antibodies enable investigation of whether IL17RC forms heterodimers with IL17RA or heterotrimers with IL17RA homodimers

  • Cross-linking studies with labeled antibodies help visualize complex formation

• Signaling pathway dissection:

  • IL17RC antibodies have revealed that the receptor associates with Act1 adaptor protein

  • They've shown that IL17 treatment leads to association with a phosphorylated form of Act1

  • Only IL17RC variants containing the SEFEX region associate with phospho-Act1

• Functional blockade experiments:

  • Neutralizing antibodies can block specific ligands (IL-17A, IL-17F, or IL-17A/F heterodimer)

  • This approach helps delineate the relative contributions of each cytokine

• Cytokine response measurements:

  • After antibody-mediated blockade, researchers can measure downstream effects on:

    • NF-κB activation

    • IL-6 and 24p3/lipocalin-2 expression

    • Production of proinflammatory mediators

Through these approaches, researchers have established that IL17RC is vital for IL-17-dependent signaling, with its extended SEFIR domain (SEFEX) required for association with phosphorylated Act1 and promotion of downstream signaling .

How can researchers differentiate between IL17RC isoforms using antibodies?

Differentiating between the numerous IL17RC isoforms requires strategic antibody selection and experimental design:

• Epitope mapping strategy:

  • Select antibodies targeting different domains of IL17RC

  • Use antibodies against constitutive regions (present in all isoforms) for general detection

  • Employ antibodies against alternatively spliced regions for isoform-specific detection

• Western blot separation:

  • Use gradient gels (4-12% or 4-15%) to maximize separation of different molecular weight isoforms

  • Run longer gels for better resolution of closely migrating variants

  • Compare migration patterns with predicted molecular weights from sequence data

• Two-dimensional electrophoresis:

  • Separate isoforms first by isoelectric point, then by molecular weight

  • Particularly useful for distinguishing post-translationally modified variants

• Deglycosylation treatments:

  • Treat samples with enzymes like PNGase F to remove N-linked glycans

  • Helps distinguish glycosylation-based size differences from actual isoform variation

  • As shown in research findings, tunicamycin treatment resolved a larger IL-17RA isoform to a single band

• Truncation isoform detection:

  • Use antibodies targeting N-terminal and C-terminal regions to identify truncated variants

  • Absence of signal with one antibody but presence with another suggests truncation

• Functional correlation:

  • Different isoforms may show distinct functional properties

  • Correlate antibody detection patterns with functional assays measuring:

    • IL-17A/F binding capacity

    • Signal transduction potential

    • Act1 phosphorylation association

The full-length isoform is estimated to occur approximately 10% of the time, while the three most common isoforms, as a group, occur about 50% of the time . Understanding this distribution helps interpret antibody detection patterns in experimental samples.

How can IL17RC neutralizing antibodies be used to study the functional relationship between IL17RC and fungal infections?

IL17RC neutralizing antibodies offer powerful tools for investigating the critical role of IL17RC in fungal defense, particularly against Candida albicans:

• In vitro neutralization assays:

  • Treat cell cultures with IL17RC neutralizing antibodies before fungal challenge

  • Measure changes in:

    • Antimicrobial peptide production (e.g., β-defensins)

    • Cytokine/chemokine expression

    • NF-κB activation

    • Fungicidal activity of cells

• Ex vivo tissue models:

  • Apply neutralizing antibodies to organotypic cultures (e.g., oral mucosa)

  • Challenge with Candida and assess invasion and tissue damage

  • Analyze epithelial cell responses and immune cell recruitment

• Comparative studies with genetic models:

  • Compare antibody neutralization effects with IL17RC knockout systems

  • Determine if antibody blockade recapitulates genetic deficiency

  • Research shows IL17RC-/- mice are susceptible to oral candidiasis similar to IL17RA-/- mice

• Neutrophil recruitment analysis:

  • Studies show IL17RC-/- mice exhibit defective neutrophil recruitment to sites of fungal infection

  • Use neutralizing antibodies to confirm this mechanism

  • Employ intravital microscopy to visualize neutrophil behavior after antibody treatment

• Temporal intervention studies:

  • Apply neutralizing antibodies at different timepoints during infection

  • Determine critical windows where IL17RC signaling is essential

  • Distinguish between roles in initial defense vs. clearance/resolution

Research demonstrates that IL17RC is vital for defense against candidiasis, with IL17RC-/- mice showing susceptibility similar to IL17RA-/- and IL-23p19-/- mice . Neutralizing antibodies offer a complementary approach to genetic models for dissecting these mechanisms.

What techniques can help resolve contradictory results when using different IL17RC antibodies?

When different IL17RC antibodies yield contradictory results, several methodological approaches can help resolve these discrepancies:

• Epitope mapping comparison:

  • Identify the target epitopes for each antibody

  • Determine if epitopes are in:

    • Different domains (extracellular vs. intracellular)

    • Constitutive vs. alternatively spliced regions

    • Native vs. denatured-only accessible regions

  • Different epitopes may explain detection of distinct subsets of IL17RC

• Isoform specificity analysis:

  • With multiple alternative splice forms of IL17RC reported , determine which isoforms each antibody detects

  • Use recombinant isoforms as positive controls

  • This may reveal that contradictory results reflect detection of different isoform subsets

• Cross-validation with knockout controls:

  • Test antibodies in IL17RC knockout systems

  • Any signal in knockout samples indicates non-specific binding

  • Helps identify false positive signals

• Post-translational modification sensitivity:

  • Test antibody detection before and after:

    • Deglycosylation (as shown with tunicamycin in research findings)

    • Dephosphorylation

    • Deubiquitination

  • May reveal modification-dependent epitope accessibility

• Orthogonal detection methods:

  • Combine antibody detection with:

    • Mass spectrometry identification

    • RNA expression correlation

    • Tagged recombinant expression

  • Convergence of multiple methods increases confidence

• Functional validation:

  • Correlate antibody detection with functional assays

  • Research shows IL17RC is required for IL-17-dependent IL-6 and 24p3 expression

  • Functional confirmation supports antibody validity

By systematically applying these approaches, researchers can determine which antibodies provide accurate results for specific applications and experimental conditions, resolving apparent contradictions.

How can IL17RC antibodies contribute to understanding the stoichiometry of the IL17RA/RC complex?

Understanding the stoichiometry of the IL17RA/RC complex requires sophisticated methodological approaches where IL17RC antibodies play crucial roles:

• Quantitative co-immunoprecipitation:

  • Use calibrated amounts of IL17RC and IL17RA antibodies

  • Immunoprecipitate receptor complexes

  • Quantify relative amounts of co-precipitated receptors

  • Analyze molar ratios to infer stoichiometry

  • Research indicates the complex "may form a heterodimer with IL-17RA, or a heterotrimer with a preexisting IL-17RA homodimer"

• Cross-linking coupled with immunoprecipitation:

  • Treat cells with membrane-impermeable or permeable cross-linkers

  • Immunoprecipitate with IL17RC antibodies

  • Analyze cross-linked products by Western blot

  • Pattern of bands indicates number of interacting partners

• Fluorescence resonance energy transfer (FRET):

  • Previous FRET studies suggest IL-17RA forms homodimers in the unliganded state

  • Label IL17RC and IL17RA with donor/acceptor fluorophore pairs via antibodies

  • Measure FRET efficiency before and after ligand addition

  • Changes indicate conformational rearrangements

• Antibody-based proximity assays:

  • Use proximity ligation assay (PLA)

  • Use antibodies against IL17RC and IL17RA

  • Signal generation requires proximity <40 nm

  • Signal intensity correlates with complex abundance

• Blue native gel electrophoresis:

  • Solubilize receptor complexes under native conditions

  • Separate by molecular weight

  • Detect with IL17RC and IL17RA antibodies

  • Determine molecular weight of intact complexes

  • Compare with theoretical weights of different stoichiometric models

These approaches can help resolve the existing uncertainty about whether IL17RC forms heterodimers with IL17RA or heterotrimers with IL17RA homodimers, as indicated in research findings . Understanding this stoichiometry is critical for developing targeted therapeutic approaches to modulate IL-17 signaling.