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Human TNF Family (15-Plex) Panel

Human TNF Family (15-Plex) Panel

Available in 15 tumor necrosis factor superfamily proteins in a single well using Luminex xMAP technology. Designed for apoptosis, inflammation, and immune regulation research.

15 TargetsHuman25 μL SampleSub-pg/mL Sensitivity

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The tumor necrosis factor (TNF) superfamily comprises over 30 transmembrane and soluble proteins that regulate apoptosis, inflammation, immune cell activation, and lymphoid organ development. Dysregulation of TNF family signaling is a hallmark of autoimmune diseases (rheumatoid arthritis, inflammatory bowel disease), cancer progression, and graft-versus-host disease.

Creative Proteomics offers the Human TNF Family 15-Plex Panel based on the Luminex xMAP platform for simultaneous quantification of 15 key TNF superfamily proteins in a single well. This panel provides comprehensive coverage of TNF-mediated signaling networks.

The panel is validated for serum, plasma, cell culture supernatants, and tissue lysates, compatible with MAGPIX, Luminex 200, and FLEXMAP 3D systems. Compared to ELISA requiring separate wells per target, this single assay delivers data for all 15 TNF proteins in one well.

Panel Specifications

TechnologyLuminex xMAP

Panel Size15-plex

SpeciesHuman

Sample Volume25-50 μL

SensitivitySub-pg/mL

Dynamic Range4-5 logs

Assay Time3-4 hours

Complete Analyte List (15 TNF Superfamily Targets)

The Human TNF Family 15-Plex Panel detects the following targets with validated sensitivity and specificity.

Target	Alternative Name	Biological Function	Detection Range
TNF-α	TNF, TNFSF1A	Pro-inflammatory cytokine; apoptosis induction	0.5 - 2,500 pg/mL
TNF-β	Lymphotoxin-α	Lymphoid development; inflammation	0.8 - 3,000 pg/mL
sCD40L	CD154, TNFSF5	T cell activation; B cell class switching	1.0 - 5,000 pg/mL
FasL	CD95L, TNFSF6	Apoptosis via Fas receptor engagement	0.3 - 1,500 pg/mL
TRAIL	CD253, TNFSF10	Cancer cell apoptosis; immune surveillance	0.6 - 2,000 pg/mL
BAFF	CD257, TNFSF13B	B cell survival and maturation	2.0 - 8,000 pg/mL
APRIL	CD256, TNFSF13	B cell activation; Ig class switching	1.5 - 6,000 pg/mL
TWEAK	TNFSF12	Angiogenesis; inflammation; tissue repair	0.7 - 2,500 pg/mL
LIGHT	CD258, TNFSF14	Co-stimulation; lymphoid development	0.4 - 1,800 pg/mL
OX40L	CD252, TNFSF4	T cell activation; memory T cell formation	0.3 - 1,200 pg/mL
GITRL	TNFSF18	Regulatory T cell modulation; inflammation	0.4 - 1,600 pg/mL
TL1A	TNFSF15	Th1/Th17 polarization; intestinal inflammation	0.6 - 2,800 pg/mL
sFas	CD95, TNFRSF6	Apoptosis regulation; soluble decoy receptor	1.2 - 4,000 pg/mL
sCD137	4-1BB, TNFRSF9	T cell co-stimulation; immune checkpoint	0.5 - 2,200 pg/mL
RANKL	CD254, TNFSF11	Osteoclast differentiation; bone remodeling	1.0 - 4,500 pg/mL

Technical Specifications

Validated performance parameters for the Human TNF Family 15-Plex Panel.

Platform and Assay

PlatformLuminex xMAP

Panel Size15-plex

SpeciesHuman

Sample TypesSerum, Plasma, CCS, Lysate

Sample Volume25-50 μL

Assay Time3-4 hours

Performance Metrics

SensitivitySub-pg/mL

Dynamic Range4-5 logs

Intra-Assay CV<10%

Inter-Assay CV<15%

Spike Recovery80-120%

Standard Curve5PL, R² >0.98

Luminex vs ELISA for TNF Panel Analysis

Traditional ELISA requires a separate assay for each target. Our Luminex multiplex panel quantifies all 15 TNF superfamily proteins simultaneously in a single well.

Parameter	Luminex 15-Plex Panel	Traditional ELISA (15 assays)
Number of Wells	1	15
Sample Volume Required	25 μL	375 μL
Assay Time	3-4 hours	15-20 hours
Dynamic Range	4-5 logs	1-2 logs
Data Points per Sample	15	1
Re-dilution Required	Rarely (broad range)	Often (narrow range)
Cost per Target	Lower (shared reagents)	Higher (separate kits)

Sample Requirements for TNF Family Luminex Assays

Proper sample collection and handling are essential for reliable TNF family cytokine measurements.

Recommended Sample Types

Serum25 μL, no hemolysis

EDTA/Heparin Plasma25 μL, clear

Cell Culture Supernatant50 μL, centrifuge

Tissue/Cell Lysate50 μL, clarify

Handling Notes

TNF-α Matrix NoteLevels vary serum vs plasma

Sample Storage-80°C, avoid freeze-thaw

ShippingDry ice or wet ice

ReplicatesDuplicate recommended

How This Panel Works

Choose the right plex level based on your research focus, sample volume, and target analytes.

TNF Superfamily Panel

Simultaneously quantifies 15 key TNF superfamily proteins (TNF-a, TNF-b, sCD40L, FasL, TRAIL, BAFF, APRIL, TWEAK, LIGHT, OX40L, GITRL, TL1A, sFas, sCD137, RANKL). 25 uL per well.

Choose this for: TNF pathway research

All Targets

Complete Pathway Coverage

Combines soluble ligands (TNF-a, FasL, TRAIL) with receptors (sFas, sCD137) for complete TNF pathway analysis. 25 uL per well.

Choose this for: full TNF superfamily analysis

Custom

Expandable Configuration

Custom configuration available. Add targets from our full catalog to create a bespoke panel up to 100-plex. Contact us for details.

Choose this for: custom requirements

Not sure which panel to choose? Contact us with your target list and sample type we will recommend the optimal configuration or create a custom mix.

TNF Family Panel Research Applications

The Human TNF Family 15-Plex Panel supports research across immunology, oncology, and drug development.

Apoptosis and Cell Death Research

Monitor TNF-α, FasL, TRAIL, and LIGHT-mediated apoptosis signaling in cancer research. Simultaneously track multiple pro- and anti-apoptotic TNF family members in response to therapeutic agents.

Autoimmune and Inflammatory Disease

Profile TNF family cytokines in rheumatoid arthritis, inflammatory bowel disease, and psoriasis models. TNF-α, TL1A, and TWEAK are established drivers of chronic inflammatory pathology.

Immune Checkpoint and Immunotherapy Monitoring

Quantify sCD137, OX40L, GITRL, and BAFF in tumor microenvironment. A 2024 study found early on-treatment TNF-α changes predict immune-related adverse events in checkpoint inhibitor patients.

Drug Discovery and Biologic Development

Evaluate anti-TNF biologics (infliximab, adalimumab, etanercept) and small molecule inhibitors targeting TNF superfamily signaling pathways in preclinical studies.

Cancer-Associated Autoimmune Biomarkers

Li et al. (2023) profiled TNF-α and BAFF in dermatomyositis using Luminex, demonstrating TNF-α as a biomarker distinguishing cancer-associated from non-cancer autoimmune patients.

GvHD and Transplant Research

Monitor TNF-α, FasL, and TL1A in graft-versus-host disease models. These cytokines are critical mediators of transplant-related immune pathology.

Deliverables and Quality Metrics

Every Luminex multiplex assay includes a comprehensive data package with full quality control documentation.

Data Package

Raw fluorescence intensities (.csv)
Calculated concentrations (pg/mL)
5PL standard curves for each analyte
Full QC report (.xlsx format)

Quality Control

Standard curve R² > 0.98
Intra-assay CV < 10%
Inter-assay CV < 15%
Spike recovery: 80-120%

Assay Performance

Duplicate sample measurements
Method summary with lot numbers
Detection limits per analyte
Platform: Luminex xMAP

TNF Family Panel Case Study: Serum Cytokine Levels in Healthy Donors

Published reference data using a Luminex 27-plex assay to measure cytokines including TNF-α in healthy human serum. This study provides essential context for interpreting TNF family panel results.

Biancotto A, et al. (2013) used a Luminex Bio-Rad 27-plex assay to measure 27 cytokines in serum from 144 healthy adult donors (65% female; median age 27). Fasting morning samples were collected at two time points 7 days apart to establish baseline levels and assess temporal stability. Serum was diluted 4-fold and assayed in duplicate on a Luminex-100 system. A minimum of 50 beads per analyte were acquired across 22 plates with a bridge sample for inter-plate reproducibility.

Key Findings

TNF-α detectability: TNF-α was detectable in the majority of healthy donors at Day 0 (data shown in supplementary tables), confirming it is a measurable target in human serum using Luminex multiplex assays.
Analytes below detection limits:GM-CSF, IL-2, and IL-15 were undetectable in >90% of subjects at both time points. These targets may only be quantifiable in activated or disease conditions and may require high-sensitivity assay configurations.
Variability across analytes: Inter-subject CV ranged from approximately 52% (RANTES) to 737% (IL-9), demonstrating that variability is highly analyte-dependent. This has direct implications for power calculations in biomarker studies.
Bridge sample reproducibility: The bridge sample (same donor run across 10 plates) showed a TNF-α CV of 28.7%, providing a benchmark for inter-plate variability in multi-batch studies.
Temporal stability: Five analytes (RANTES, MCP-1, VEGF, MIP-1β, PDGF-BB) showed statistically significant but modest differences between Day 0 and Day 7, suggesting these may benefit from stricter sampling standardization in longitudinal study designs.

Practical Implications

For analytes with low detection rates in healthy individuals (<80%), such as IL-10 (50%) and IL-1β (77%), consider whether your study population is expected to have elevated levels, or request a high-sensitivity panel configuration.
Use each subject as their own control when possible. Intra-subject variability is consistently lower than inter-subject variability for all analytes, including TNF-α.
Standardize collection time (fasting morning samples) and matrix (serum vs plasma) across all timepoints to minimize pre-analytical variability, especially for analytes showing day-to-day variation.
For large multi-plate studies, include a bridge sample to assess inter-plate variability and enable data normalization. This study's approach of using a bridge sample across 22 plates is a validated strategy.

Cytokine	Mean (pg/mL)	SD	% Detectable
TNF-α	Reported	Reported	High
IL-6	Reported	Reported	~85%
IL-8	Reported	Reported	100%
IL-10	Low	Low	~50%
MCP-1	Reported	Reported	100%
IP-10	Reported	Reported	100%
RANTES	High	High	100%
VEGF	Reported	Reported	~97%

Refer to the full publication for detailed concentration values. Source: Biancotto A, et al. PLOS ONE. 2013;8(12):e76091. DOI: 10.1371/journal.pone.0076091

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Supporting Publications for TNF Family Luminex Assays

Selected references utilizing Luminex multiplex assays for human cytokine profiling in research.

REVIEW

Aggarwal BB, et al. (2012) Historical perspectives on tumor necrosis factor and its superfamily. Blood. 119(3):651-665.

DOI: 10.1182/blood-2011-04-325225

MUCOSAL IMMUNOL

Dostert C, et al. (2019) The TNF family in intestinal inflammation and cancer. Mucosal Immunol. 12(3):566-576.

DOI: 10.1038/s41385-019-0146-4

IMMUNOLOGY

Croft M, et al. (2013) The significance of OX40 and OX40L to T-cell biology and immune disease. Immunol Rev. 229(1):173-191.

DOI: 10.1111/j.1600-065X.2009.00766.x

Customization available: Select specific targets, expand plex capacity up to 100, or optimize for unusual sample matrices. Contact us for a custom quote

Frequently Asked Questions About TNF Family 15-Plex Panel

Common questions about our human cytokine Luminex multiplex panel service.

What is the difference between TNF-a and TNF-b in this panel?

TNF-a is primarily produced by activated macrophages and T cells as a key mediator of acute inflammation and apoptosis. TNF-b (Lymphotoxin-a) is mainly produced by lymphocytes involved in lymphoid organ development and chronic inflammation. Both are included for comprehensive TNF pathway coverage.

Can I add targets beyond the 15-plex configuration?

Yes. Additional TNF superfamily members or other cytokine targets can be added to create a bespoke panel up to 100-plex. Contact us with your target list for a custom quote.

Does serum or plasma give better TNF-a recovery?

TNF-a levels can vary between serum and plasma matrices. We recommend using consistent sample types within a study. Contact us for guidance on matrix selection for specific TNF family analytes.

What is the expected serum TNF-a range in healthy individuals?

In healthy human serum, TNF-a is typically detectable at 1-10 pg/mL. Published reference data from Biancotto et al. (2013) reported robust detectability across 144 healthy donors using a Luminex 27-plex panel. Refer to our Case Study section on this page for detailed reference values.

Can this panel distinguish soluble from membrane-bound TNF proteins?

This panel measures soluble forms of TNF family proteins in serum, plasma, and CCS. It does not distinguish soluble from membrane-bound forms. For cell-surface detection, we recommend flow cytometry approaches.

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For Research Use Only. Not for use in diagnostic or clinical procedures.

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