LUMINEX MULTIPLEX ASSAY SERVICE

Rat Th1/Th2/Th17 Cytokine Multiplex Panel (14-Plex)

Simultaneous quantification of 14 key rat T helper cytokines — covering Th1, Th2, and Th17 axes plus myeloid growth factors — in a single well using Luminex xMAP technology. Designed for comprehensive preclinical T cell immunology, vaccine studies, and immune response profiling in rat models.

14 TargetsRat25 μL Samplepg/mL Sensitivity
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14-Plex Th1/Th2/Th17
Brown University
Harvard University
Imperial College London
University of Florida
Tulane University
Abata Therapeutics
AlzeCure Pharma

T helper (Th) cells are the orchestrators of adaptive immunity, directing the character of the immune response through the cytokines they secrete. Th1 cells produce IFN-γ, IL-2, and TNF-α to drive cell-mediated immunity against intracellular pathogens and tumors. Th2 cells produce IL-4, IL-5, and IL-13 to drive humoral immunity against extracellular parasites and allergens. Th17 cells produce IL-17A to drive neutrophil-mediated inflammation and mucosal antifungal defense. The balance among these T helper subsets — Th1/Th2 ratio and Th17 axis — determines the character of the immune response, and its dysregulation underlies autoimmune disease, allergy, and impaired anti-tumor immunity.

Creative Proteomics offers the Rat Th1/Th2/Th17 Cytokine 14-Plex Panel based on the Luminex xMAP platform for simultaneous quantification of 14 key T helper cytokines in a single well. The panel covers Th1 cytokines (IFN-γ, IL-2, IL-12p70, TNF-α), Th2 cytokines (IL-4, IL-5, IL-10, IL-13), Th17 effector (IL-17A), pro-inflammatory mediators (IL-1α, IL-1β, IL-6), and myeloid growth factors (GM-CSF, G-CSF) — providing complete coverage of the T helper polarization spectrum in a single well. The panel is available as a focused Th1/Th2 12-plex subset or the full 14-plex. Validated for serum, plasma, and cell culture supernatants, the panel is compatible with MAGPIX, Luminex 200, and FLEXMAP 3D systems.

The panel is backed by validated performance data with intra-assay CV <10% and broad dynamic range, enabling accurate quantitation from a single 25 μL sample. It includes the full analyte set of the widely cited Th panel format used in hundreds of published rat immunology studies, plus G-CSF for myeloid activation context.

Panel Specifications
TechnologyLuminex xMAP
Panel Size14-plex (Th1/Th2 12-plex subset also available)
SpeciesRat
Sample TypesSerum, EDTA/Heparin Plasma, CCS
Sample Volume25 μL per well
Sensitivitypg/mL (varies by analyte)
Assay Time~4 hours

Complete Analyte List — 14 Rat Th1/Th2/Th17 Cytokines

The Rat Th1/Th2/Th17 14-Plex Panel covers the full Th1/Th2/Th17 spectrum plus key pro-inflammatory mediators and myeloid growth factors. The 12-plex Th1/Th2 subset (excluding IL-17A and G-CSF) is also available.

Target Alternative Name T Helper Axis Biological Function
IFN-γ Interferon-gamma, Type II IFN Th1 Signature Th1 cytokine; activates macrophages for enhanced microbicidal activity and antigen presentation; upregulates MHC class I and II; promotes IgG2a class switching in rodents; elevated in autoimmune and infectious disease models
IL-2 Interleukin-2, T Cell Growth Factor Th1 / Treg Primary T cell growth factor driving clonal expansion of activated T cells; also essential for regulatory T cell (Treg) maintenance and peripheral tolerance; IL-2/IL-2R axis is a therapeutic target in transplantation and autoimmunity
IL-12p70 Interleukin-12 p70 Th1 Polarization Heterodimeric cytokine (p35 + p40 subunits); master inducer of Th1 differentiation; stimulates IFN-γ production from NK and T cells; bridges innate sensing to adaptive immunity; p40 subunit shared with IL-23
TNF-α Tumor Necrosis Factor-alpha, Cachectin Th1 / Pro-Inflammatory Early-response cytokine produced by activated macrophages and Th1 cells; drives endothelial activation, vascular permeability, and neutrophil recruitment; key therapeutic target in RA, IBD, and psoriasis models
IL-4 Interleukin-4, B Cell Stimulatory Factor-1 Th2 Signature Th2 cytokine; drives naive CD4+ T cell differentiation toward Th2 phenotype; promotes B cell class switching to IgG1 and IgE; suppresses Th1 differentiation; central to allergic and anti-parasitic responses
IL-5 Interleukin-5, Eosinophil Differentiation Factor Th2 Eosinophil growth, differentiation, and activation factor; promotes IgA production by B cells; elevated in allergic airway inflammation and parasitic helminth infection models
IL-10 Interleukin-10, CSIF Th2 / Treg Prototypic anti-inflammatory cytokine; suppresses macrophage and dendritic cell function; inhibits IL-12, TNF-α, and IFN-γ production; essential for immune homeostasis and resolution of inflammation
IL-13 Interleukin-13 Th2 Shares IL-4Rα signaling chain with IL-4; drives goblet cell metaplasia, mucus hypersecretion, and airway hyperresponsiveness; central effector in allergic asthma models; promotes collagen deposition and fibrosis
IL-17A Interleukin-17A, CTLA-8 Th17 Signature Th17 cytokine; induces neutrophil-recruiting chemokines (CXCL1, CXCL8), G-CSF, and antimicrobial peptides from epithelial and stromal cells; critical for mucosal antifungal defense and barrier immunity; pathogenic in psoriasis, RA, EAE, and IBD models
GM-CSF Granulocyte-Macrophage Colony-Stimulating Factor T Cell / Myeloid Growth factor for granulocyte and macrophage lineages; produced by activated T cells; promotes dendritic cell maturation and antigen presentation; links T cell activation to myeloid effector recruitment
G-CSF Granulocyte Colony-Stimulating Factor, CSF-3 Myeloid Growth Factor Primary regulator of neutrophil production and mobilization from bone marrow; induced by IL-17A in stromal and epithelial cells; elevated in bacterial infection, sterile inflammation, and Th17-driven pathology; provides myeloid context to the T helper cytokine profile
IL-1α Interleukin-1 alpha, IL-1F1 Pro-Inflammatory Constitutively expressed alarmin released upon cell damage/necrosis; activates IL-1R1 to initiate sterile inflammation; unlike IL-1β, does not require inflammasome processing for activity
IL-1β Interleukin-1 beta, IL-1F2 Pro-Inflammatory Prototypic inflammasome-dependent cytokine; cleaved by caspase-1 following NLRP3/AIM2/NLRC4 activation; potent pyrogen driving fever and acute-phase protein synthesis
IL-6 Interleukin-6, BSF-2 Pro-Inflammatory / Th17 Master coordinator of the acute-phase response; induces hepatic CRP and fibrinogen; drives Th17 differentiation in combination with TGF-β1; promotes B cell antibody secretion
Th1/Th2/Th17 balance in rodent models: In rats, the three T helper axes drive distinct immune effector mechanisms: Th1 (IFN-γ, IL-12) drives cell-mediated immunity and IgG2a class switching; Th2 (IL-4, IL-5, IL-13) drives humoral immunity and IgG1/IgE; Th17 (IL-17A) drives neutrophil-mediated inflammation and mucosal defense. The 14-plex panel captures all three axes simultaneously, plus G-CSF as the key downstream effector of IL-17A signaling and GM-CSF as the T cell-to-myeloid bridge.

Need the Th1/Th2 12-plex subset or broader coverage? This 14-plex can be configured as the focused 12-plex (Th1/Th2 only, excluding IL-17A and G-CSF), or expanded with additional chemokines (MCP-1, RANTES, IP-10) and growth factors (VEGF). Contact us for your preferred configuration.

Technical Specifications

Validated performance parameters for the Rat Th1/Th2/Th17 Cytokine 14-Plex Panel. The 12-plex Th1/Th2 subset shares identical specifications.

Platform and Assay
PlatformLuminex xMAP (MAGPIX / Luminex 200 / FLEXMAP 3D)
Panel Size14-plex (12-plex Th1/Th2 subset also available)
SpeciesRat
Sample TypesSerum, EDTA/Heparin Plasma, CCS
Sample Volume25 μL per well
Sample DilutionNeat to 1:2 (serum/plasma)
Assay Time~4 hours
Performance Metrics
Sensitivitypg/mL range (varies by analyte)
Intra-Assay CV<10%
Inter-Assay CV<15%
Spike Recovery80–120%
Cross-ReactivityNegligible between antibodies within the panel
Bead TypeMagnetic MagPlex microspheres

Luminex Multiplex vs ELISA for Rat T Helper Cytokine Analysis

T helper polarization is defined by the relative levels of multiple cytokines across Th1, Th2, and Th17 axes. Luminex multiplex captures the full Th polarization profile from a single sample.

Parameter Luminex 14-Plex Traditional ELISA (14 assays)
Targets per Well 14 1
Sample Volume 25 μL 350–700 μL total
Assay Time ~4 hours 42–56 hours total
Th Polarization Insight Th1/Th2 ratio + Th17 axis from same aliquot Requires 3+ separate assays; between-assay CV invalidates ratios
Comprehensive Profiling All 14 cytokines from one 96-well plate 14 plates with different standard curves and QC

The IFN-γ/IL-4 ratio remains the gold standard index of Th1/Th2 balance in rodent immunology. The IL-17A measurement adds the critical Th17 dimension — elevated IL-17A with normal IL-4 distinguishes Th17-driven autoimmune pathology (psoriasis, EAE) from Th2-driven allergic disease, with direct implications for therapeutic strategy (anti-IL-17 vs. anti-IL-4/IL-13). The 14-plex panel captures all three T helper axes from a single 25 μL sample, eliminating the between-assay variability that would invalidate cross-axis comparisons if measured by separate ELISAs.

Sample Requirements for Rat T Helper Cytokine Luminex Assays

Proper sample collection and handling are critical for accurate T helper cytokine measurement. Th2 and Th17 cytokines (IL-4, IL-5, IL-13, IL-17A) are typically at lower concentrations than Th1 cytokines and require careful handling.

Sample Type Volume Requirement
Serum 25 μL Collect in SST tubes; allow clotting for 30 min at room temperature; centrifuge at 1,500g for 10 min. Serial sampling via tail vein for longitudinal Th polarization studies.
EDTA/Heparin Plasma 25 μL Centrifuge within 30 min at 2,500g for 15 min; platelet-poor plasma recommended to minimize platelet-derived cytokines (TNF-α, IL-1β)
Splenocyte Culture Supernatant 50 μL Stimulate with PMA/ionomycin or anti-CD3/CD28 for 24–72 hours; centrifuge at 10,000g for 10 min; the gold standard for ex vivo Th1/Th2 polarization assessment
Lymph Node Cell Supernatant 50 μL Same stimulation protocol as splenocytes; draining lymph nodes preferred for antigen-specific T cell responses
Minimum Project Size — One 96-well plate; smaller batches accepted with surcharge
Sample Storage — -80°C; avoid repeated freeze-thaw (>2 cycles may degrade IL-4 and IL-13 disproportionately vs. IFN-γ, artifactually skewing Th1/Th2 ratios)
Shipping — Dry ice; samples must remain frozen throughout transit

How the Rat Th1/Th2/Th17 14-Plex Panel Works

The T helper response is not binary — it exists on a Th1-Th2-Th17 continuum with myeloid context. The 14-plex panel provides four layers of information about the T helper polarization state.

Th1

Th1 Axis

IFN-γ, IL-2, IL-12p70, TNF-α — cell-mediated immunity driven by Th1-polarized CD4+ T cells. IFN-γ is the signature cytokine activating macrophages. IL-12p70 is the master inducer driving naive T cells toward Th1. 25 μL per well.

Key readout: IFN-γ, IFN-γ/IL-4 ratio
Th2

Th2 Axis

IL-4, IL-5, IL-10, IL-13 — humoral immunity and allergic inflammation. IL-4 is the master inducer of Th2 differentiation. IL-5 drives eosinophilia. IL-13 drives mucus production and fibrosis. 25 μL per well.

Key readout: IL-4, IL-5, IL-13
Th17

Th17 Axis & Myeloid Link

IL-17A, IL-6, G-CSF, GM-CSF — IL-17A is the signature Th17 cytokine driving neutrophil-mediated inflammation. G-CSF is the key downstream effector induced by IL-17A in stromal cells. IL-6 (with TGF-β1) drives Th17 differentiation. 25 μL per well.

Key readout: IL-17A, G-CSF, IL-17A/IFN-γ ratio
Why three axes matter: In the rat EAE model, disease was historically attributed to Th1 (IFN-γ). It is now understood that IL-17A-producing Th17 cells are the primary pathogenic drivers, with IFN-γ playing a regulatory role. Anti-IL-17 therapy is effective in EAE, while anti-IFN-γ exacerbates disease. A panel measuring only Th1/Th2 would miss the dominant pathogenic axis. The 14-plex captures all three axes — Th1, Th2, and Th17 — plus G-CSF as the downstream myeloid amplifier, providing the complete T helper polarization profile.

Rat Th1/Th2/Th17 Cytokine Panel Research Applications

The Rat Th1/Th2/Th17 14-Plex Panel supports preclinical research across T cell immunology, vaccine development, autoimmunity, and infectious disease.

Autoimmune Disease Models

Rat models of rheumatoid arthritis (CIA), multiple sclerosis (EAE), and type 1 diabetes (BB rat, STZ) are characterized by Th1/Th17-skewed cytokine profiles. The 14-plex panel quantifies the Th1 (IFN-γ, IL-2, TNF-α) and Th2 (IL-4, IL-10) balance at baseline, during disease induction, and in response to immunomodulatory therapy from serial serum or splenocyte samples.

Vaccine Adjuvant and Immunogenicity Studies

Vaccine adjuvants are classified by their ability to polarize T helper responses: alum drives Th2 (IL-4, IL-5 dominant), CpG and poly I:C drive Th1 (IFN-γ, IL-12 dominant). The 12-plex panel profiles the Th1/Th2 cytokine signature induced by candidate adjuvants and vaccine formulations in rat models, providing quantitative, multiplexed readouts of T helper polarization.

Allergy and Asthma Models

Ovalbumin (OVA)-induced allergic airway inflammation in Brown Norway rats is the classic Th2-driven model. IL-4, IL-5, and IL-13 are the central effector cytokines driving IgE production, eosinophil recruitment, and airway hyperresponsiveness. The panel quantifies all three Th2 cytokines alongside Th1 counter-regulators (IFN-γ) from BAL fluid, serum, and lung homogenates.

Transplantation Immunology

Acute allograft rejection is Th1-driven (IFN-γ, IL-2 dominant), while Th2 polarization (IL-4, IL-10) is associated with graft acceptance and tolerance. The panel enables serial monitoring of the Th1/Th2 balance in rat models of heart, kidney, and liver transplantation to assess immunosuppressive regimen efficacy and tolerance induction.

Infectious Disease and Host Defense

Intracellular pathogens (Listeria, Toxoplasma) require Th1 responses for clearance (IFN-γ-mediated macrophage activation). Helminth infections (Nippostrongylus, Trichinella) drive Th2 responses (IL-4, IL-5, IL-13 for IgE and eosinophil-mediated expulsion). The 12-plex panel distinguishes Th1-adequate from Th1-deficient responses in infection models.

Immunotoxicology and Safety Assessment

Drug-induced immunomodulation is a key safety concern in preclinical development. The 12-plex panel provides a standardized Th1/Th2 cytokine profile for immunotoxicity assessment — detecting unintended Th1 suppression (increased infection risk) or Th2 skewing (allergy/autoimmunity risk) in response to candidate therapeutic compounds.

Deliverables and Quality Metrics

Every Luminex rat Th1/Th2 cytokine assay includes a comprehensive data package with full quality control documentation.

Data Package
  • Raw fluorescence intensities (.csv)
  • Calculated concentrations (pg/mL) for all 12 cytokines
  • Th1/Th2 ratio: IFN-γ/IL-4 (optional)
  • 5PL standard curves for each analyte (R² >0.99)
  • Full QC report (.xlsx format)
Quality Control
  • Standard curve: 7-point dilution series, 5PL fit, R² >0.99
  • Intra-assay CV <10%
  • Inter-assay CV <15%
  • Spike recovery: 80–120%
Assay Performance
  • Duplicate sample measurements for all samples
  • Method summary with reagent lot numbers
  • LLOQ reported per analyte
  • Platform: Luminex xMAP, compatible with MAGPIX, Luminex 200, FLEXMAP 3D

Related Panels

Explore other Luminex panels available for rat immunology and preclinical research.

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← All Rat PanelsBrowse rat species overview

Frequently Asked Questions About Rat Th1/Th2/Th17 Panel

Common questions about our rat Th complete cytokine Luminex multiplex panel service.

How does the 14-plex differ from the standard Th1/Th2 12-plex?
The 14-plex adds two critical analytes beyond the standard Th1/Th2 12-plex: IL-17A (signature Th17 cytokine) and G-CSF (myeloid growth factor, downstream of IL-17A). These additions provide Th17 axis coverage and myeloid activation context that are absent from standard Th1/Th2 panels. The 12-plex subset is available if your research is specifically focused on Th1/Th2 polarization only.
Why is the IFN-γ/IL-4 ratio important in rat immunology?
The IFN-γ/IL-4 ratio is the most widely used index of Th1/Th2 balance in rodent models. IFN-γ is the prototypic Th1 cytokine (cell-mediated immunity); IL-4 is the prototypic Th2 cytokine (humoral/allergic immunity). A high ratio indicates Th1 dominance (autoimmunity, transplant rejection, intracellular infection). A low ratio indicates Th2 dominance (allergy, helminth infection, tolerance). The ratio is only valid when both cytokines are measured simultaneously from the same sample aliquot — separate ELISA measurements introduce between-assay variability that can obscure or artifactually create ratio differences.
Should I measure cytokines from serum or from stimulated splenocytes?
Both are informative but answer different questions. Serum/plasma cytokine levels reflect the systemic inflammatory state at the moment of blood collection — useful for monitoring disease progression or treatment response in vivo. Stimulated splenocyte or lymph node cell supernatants reveal the T cell polarization potential — what cytokines the T cells are capable of producing when activated. For comprehensive immunophenotyping, we recommend collecting both: serum for in vivo cytokine levels and splenocyte supernatants (PMA/ionomycin, 24–48 hr) for ex vivo T cell polarization capacity.
Does the 14-plex include Th17 coverage?
Yes. The 14-plex includes IL-17A, the signature Th17 effector cytokine, alongside IL-6 (which drives Th17 differentiation) and G-CSF (the key downstream effector induced by IL-17A in stromal cells). This provides complete Th17 axis coverage that standard Th1/Th2 panels lack. If you only need Th1/Th2 data, the 12-plex subset (excluding IL-17A and G-CSF) is available at a reduced configuration.
Why are IL-4, IL-5, and IL-13 often harder to detect than IFN-γ?
Th2 cytokines (IL-4, IL-5, IL-13) are typically produced at lower levels than Th1 cytokines (IFN-γ, TNF-α) in most rat models, and they are more susceptible to degradation during sample handling. IL-4 in particular has a short half-life and can be consumed by target cells expressing the IL-4 receptor. For reliable Th2 cytokine measurement: process samples quickly, avoid repeated freeze-thaw, and consider using stimulated cell culture supernatants (which concentrate the cytokines) rather than serum for Th2-dominant models like allergy or helminth infection.
Can this panel be used for non-rat rodent species?
The panel is validated and optimized for rat cytokines. While some antibodies may cross-react with mouse cytokines due to sequence homology, the standard curves and validation are rat-specific. For mouse Th1/Th2 studies, we recommend our Mouse Th Cytokine Panel which uses mouse-specific capture and detection antibodies. For other rodent species (gerbil, hamster, guinea pig), contact us to discuss cross-reactivity testing or custom assay development.

Interested in Rat T Helper Cytokine Profiling?

Contact us to discuss your rat immunology study requirements, panel configuration (14-plex Th1/Th2/Th17 or 12-plex Th1/Th2 subset), and sample collection protocols. We respond within 24 hours.

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