Introduction: Why Study Mouse Cytokines with Olink

Many preclinical studies hit a wall when assessing immune responses: low sample volume, high variability, and poor sensitivity make it hard to see subtle changes in cytokine levels. That slows discovery, especially when modeling inflammation, immuno-oncology, or vaccine response.

Using Olink's Target 48 Mouse Cytokine Panel via Creative Proteomics solves many of these issues. With just 1 µL of mouse plasma or serum, researchers can measure ≈43 validated cytokines and immune mediators with high sensitivity.

In this guide, you will gain a practical, detailed overview of how this panel works, its advantages, how it stacks up against alternative technologies, and how to integrate it into your own workflows. Whether you're doing longitudinal mouse model studies, working on immunomodulatory drug development, or probing immune pathways, this panel can make cytokine profiling more accurate, efficient, and reproducible.

What Is the Olink Target 48 Mouse Cytokine Panel?

The Olink Target 48 Mouse Cytokine Panel is a multiplex protein biomarker assay crafted for non-clinical, preclinical research. It enables measurement of ~43 immune-related proteins (cytokines, chemokines, growth factors, immune modulators) in mouse samples using just 1 µL of plasma, serum or other compatible sample types.

Here are its defining features:

Key Technical Details

Number of proteins: ~43 immune mediators.

Sample volume requirement: 1 µL of mouse plasma or serum.

Readout formats:

Absolute quantification in standard concentration units (pg/mL), using built-in calibrators.
Relative quantification in NPX (Normalized Protein Expression) units, if desired.

Sample Types & Compatibility

Primary: mouse plasma and serum.

The assay is validated for use with other matrices: e.g. citrate plasma, heparin plasma. Some additional sample types are compatible (with caveats for interference).

Performance & Validation Highlights

Sensitivity & specificity: Carefully validated; selection of biomarkers is based on relevance to mouse immune biology and minimal cross-reactivity.

Dynamic range: For various proteins, the panel has a broad measurable range (LOD → ULOQ) of several orders of magnitude. For example, for IL-3, the LOD is ~0.23 pg/mL and the ULOQ ~35,452 pg/mL.

Precision (repeatability and reproducibility): Within-run and between-run coefficient of variation (CV) are low (often in single digits), indicating reliable performance.

What It Enables Researchers to Do

Monitor multiple cytokines simultaneously in small-volume mouse samples—helpful for longitudinal studies (multiple time points from same animals).
Detect subtle immune changes in disease or experimental models (e.g., inflammation, immune stimulation, immuno-oncology) due to its high sensitivity.
Reduce sample consumption and animal numbers, because fewer animals are needed when one can collect multiple time-points or smaller volumes.

Olink Mouse Cytokine Panel workflow and features infographic Figure 1. Workflow and key features of the Olink Target 48 Mouse Cytokine Panel, highlighting sample requirements, detection method, and applications.

Types and Comparison: Mouse-Specific vs Human / Other Olink Panel Options

When selecting a panel, understanding what exists is critical. Below I compare the Olink Target 48 Mouse Cytokine Panel with other Olink offerings, especially human panels, plus some alternative technologies. This helps you choose what best fits your study.

Olink Panel Types at a Glance

Panel	Species / Scope	Number of Proteins	Sample Volume	Readout Format	Typical Use-Cases
Target 48 Mouse Cytokine	Mouse only	~43 immune mediators (cytokines, chemokines, growth factors)	1 µL plasma or serum	Absolute quantification (pg/mL) or relative units	Longitudinal mouse model studies, immuno-oncology, inflammation modeling, translational preclinical work
Target 48 Human Cytokine / Immune Surveillance	Human	~45 immune-related proteins (cytokines etc.)	1 µL human plasma / serum	Absolute (pg/mL) or relative (NPX) units	Profiling immune state, biomarker discovery in human samples, comparing human cohorts, etc.
Target 96 / Human-Targeted Panels	Primarily human (one exploratory mouse panel exists in some "Target 96 Mouse Exploratory")	~92 proteins per panel for human panels; "exploratory" mouse panel in one Target 96 as reported	Still low sample volume (~1 µL) for many panels; depending on number of proteins, sometimes more volume needed or segmented workflow	Usually absolute quantification; some relative units possible for human panels	When you need broader biomarker coverage in human samples, hypothesis generation, capturing multiple pathways etc.
Other Platforms (Non-Olink)	Varies (human, mouse)	Often fewer proteins, sometimes multiplex bead assays, ELISAs etc.	Sample volumes often larger; more hands-on prep	Absolute concentrations (some), but dynamic range, sensitivity, cross-reactivity may vary greatly	Good for targeted validation, but may be less efficient for large panels or small sample volumes

Strengths of the Mouse-Specific Panel vs Human Panels and Alternatives

Species specificity: Mouse panel is tailored for murine immune biology. Human panels have proteins optimized for human expression; cross-reactivity isn't guaranteed. Using the mouse panel ensures you're measuring the right isoforms and expression patterns.

Low sample volume: Both human and mouse Target 48 panels require ~1 µL of sample, which enables repeated sampling (longitudinal studies) in mice without sacrificing animal welfare.

Validated performance: The mouse panel shows good precision and sensitivity, with well-defined detection limits for many mouse cytokines (e.g., IL-2 has LOD ~0.26 pg/mL, LLOQ ~0.77 pg/mL).

Pathway coverage: The 43 proteins in the mouse cytokine panel span multiple immune pathways (Th1, Th2, Th17, cytokine-receptor interactions, etc.), allowing broad immune profiling.

Limitations & Trade-Offs

Breadth vs depth: With ~43 targets, the mouse panel is mid-plex. If your study requires hundreds of proteins or discovery of unexpected biomarkers outside cytokines/immune mediators, high-plex panels (e.g. Explore HT) may be more suitable.

Human vs translational comparison caveats: If you wish to compare directly between mouse and human data, using human panels may give more direct comparability—but species differences remain. The mouse panel can help, but target overlaps and expression patterns differ.

Cost and throughput: Higher-plex panels often cost more per sample and have more complex data analysis. With more proteins, you may need to consider missing data, dynamic range for low-abundance proteins, and QC.

Matrix effects and background: Even with PEA technology, sample type (plasma, serum vs other matrices), hemolysis, lipemia, etc., can affect results. These effects may differ between species or human vs mouse samples.

Practical Guidance: Which Panel to Use When

Use Target 48 Mouse Cytokine when your work is entirely in mouse models: immune modulation, treatment effect, disease modeling. Small sample sizes or repeated measures favor this panel.
Use Target 48 Human Cytokine / Immune Surveillance when working with human samples or when comparing human vs mouse responses, or for translational bridging.
Consider Target 96 human (or exploratory) panels if you suspect many pathways may be involved and you want to capture a broader proteomic signature.
Use non-Olink multiplex or ELISA methods for targeted validation, especially when only a few cytokines are of interest, or for very low-budget setups.

Advantages of Olink's Mouse Panel

Olink's Target 48 Mouse Cytokine Panel delivers several advantages that address common bottlenecks in non-clinical immune research using mouse models. Below are the key benefits supported by current validation data.

Minimal Sample Volume & Longitudinal Design

Requires only 1 µL of mouse plasma or serum per sample. This is especially useful in longitudinal studies where repeated sampling from the same animal is required.

Smaller sample volume means fewer animals needed, as multiple timepoints can be gathered from the same subject. This supports ethical research practices and reduces biological variability.

High Sensitivity, Specificity, and Broad Dynamic Range

The panel has been validated for high specificity (low cross-reactivity), meaning each antibody pair largely responds only to its intended target. Validation data show minimal cross-signal among high-homology targets.

It supports both absolute quantification (e.g. pg/mL) and relative units, enabling flexible experimental design and better cross-study comparability.

The dynamic range covers low-abundance cytokines through to high levels relevant to strong immune responses. The validation documents show measurable ranges, inter- and intra-run variation (CVs) generally in acceptable ranges (single digits to low double digits percent) for many analytes.

Streamlined Workflow & Data Reliability

Use of Proximity Extension Assay (PEA) technology: dual antibody recognition plus DNA tags ensures that only properly bound antibody pairs generate signal; this reduces background noise and improves specificity.

Built-in internal and external controls, calibrators, and quality control checks. This ensures measurement consistency and helps detect and correct for sample handling or technical variation.

Compatibility with Olink's Signature Q100 instrument and standard workflows helps reduce hands-on time. This supports more consistent throughput and reproducibility.

Applications in Preclinical and Translational Research

Olink's Target 48 Mouse Cytokine Panel offers powerful utility in many non-clinical research settings. Below are some of the best use-cases, with real data and examples, that illustrate what this panel enables in practice.

Key Preclinical Use-Cases

Longitudinal Studies in Mouse Models

Because the panel needs only 1 µL of plasma or serum, you can repeatedly sample from the same mouse over time. This reduces inter-animal variability and boosts statistical power.

Immune Response & Inflammation Research

The 43-protein panel covers many cytokines, chemokines, and immune mediators relevant to inflammation, allowing detailed monitoring of immune signaling pathways. This helps researchers dissect how treatments modulate innate or adaptive immunity.

Immuno-Oncology Models

In models of cancer, the panel enables detection of multiple immune checkpoints, growth factors, and chemotactic molecules, which helps in profiling the tumor microenvironment, immune infiltration, or response to immunomodulatory agents.

Disease Modeling & Translational Bridging

From metabolic regulation, neuroinflammation, to autoimmune-like phenotypes, the panel provides data that can bridge between mouse models and hypotheses relevant to human biology. For example, Olink reports its use in obesity, asthma, Alzheimer's disease models, showing relevance of specific cytokine changes.

Example Data from Literature / Validation

The validation documentation shows intra- and inter-run coefficient of variation (CV) across multiple proteins is acceptable, ensuring reproducibility across operators and plates.

The assay was tested on mice of various strains (e.g. C57BL/6, BALB/c, DBA/1) and models including allergen challenge, metabolic disease, autoimmune models, showing the panel can detect cytokine differences in genetically and experimentally diverse settings.

Translational Research Benefits

Hypothesis generation: Because of the panel's breadth, researchers can find unexpected cytokine changes that suggest new mechanistic insights or therapeutic targets.

Reduced animal usage: Longitudinal sampling means fewer animals needed for time-course studies.

Better comparability: Data in absolute units (pg/mL) allow better cross-study or cross-species comparisons when using similar panels in human or other systems.

Workflow integration: Compatible with the Signature Q100 instrument, Olink provides streamlined data output, QC tools, and validation that supports rigorous research.

Data Quality, Interpretation, and Workflow for Mouse Studies

Getting reliable data from mouse cytokine panels isn't just about running the assay. It requires careful workflow design, quality controls, and clear interpretation. Below are best practices, performance metrics, and what to watch out for when using the Olink Target 48 Mouse Cytokine Panel.

Workflow & Sample Preparation

Always start with 1 µL of mouse sample (plasma or serum) for each assay, as recommended.

Use compatible sample types. Plasma and serum are validated. Some other matrices (e.g. citrate or heparin plasma) are also compatible, but may need specific validation.

Handle sample collection, storage, and thawing carefully. Freeze-thaw cycles, hemolysis, lipemia, or other sample quality issues can affect low abundance cytokines. Effects are protein-dependent.

Internal Controls, Calibration & QC Checks

The panel uses internal calibrators and controls to track performance of each sample. These typically include Sample Control, Negative Control, and a Calibrator spike. These allow detection of outliers and plate-to-plate variation.

For example, Eotaxin assay has repeatability (within-run CV) ~4%, between-run CV ~3%.

Accuracy and dynamic range: each assay has a lower limit of quantification (LLOQ), upper limit (ULOQ), and limit of detection (LOD). For IL-2: LOD ~0.260 pg/mL; LLOQ ~0.770 pg/mL; ULOQ ~9,250 pg/mL.

Precision, Reproducibility & Validation Data

Olink's validation data show low intra- and inter-run variation across multiple operators. For the mouse cytokine panel, multiple plate runs (three operators, thirteen plates) show consistent CVs.

Measured background / "normal" expression in control mice shows distribution for many cytokines. This gives benchmarks for expected ranges in healthy vs experimental models.

Interpretation: Units, Normalization, and Data Analysis

Results are delivered either as absolute concentrations (pg/mL) via calibrator curves, or relative units (Normalized Protein Expression, NPX) depending on experimental needs. The absolute quantitation helps cross-study comparison; NPX can help with relative changes.

Be mindful of lower abundance cytokines: if measured values are close to LOD / LLOQ, statistical confidence is lower. Consider replicates or validation via orthogonal methods for low signal proteins.

Normalize data carefully: sample handling, matrix effects, and interfering substances (hemolysis, etc.) can add bias. Use built-in QC flags and controls to filter out poor-quality samples.

How to Integrate Olink's Mouse Panel Into Your Workflow

Integrating Olink's Target 48 Mouse Cytokine Panel into your lab's research workflow involves planning around sample prep, QC, instrument use, data analysis, and logistics. Below are best practices grounded in Olink's published validation data and product specifications.

Step-by-Step Integration Checklist

Step	Key Actions	Why It's Important
1. Experimental Design	Define time points/longevity (longitudinal vs terminal study) Choose number of animals per group Determine strain, age, sex of mice (strain background affects baseline cytokine levels)	Ensures statistical power, reduces inter-animal variability, improves interpretability.
2. Sample Collection & Handling	Collect plasma or serum under consistent conditions (same anticoagulant, bleed site) Minimize hemolysis, lipemia Aliquot and freeze at −80 °C Avoid repeated freeze-thaw cycles	Uniform sample handling preserves low-abundance cytokine integrity.
3. Sample Volume & Compatibility	Use only 1 µL of sample per assay as specified for the mouse panel. Confirm sample type (plasma, serum, or other validated matrices) is accepted.	Conserves precious samples; validated performance depends on using approved matrices.
4. Assay & Plate Setup	Use Signature Q100 instrument (as compatible with the mouse panel). Include internal controls, calibrators, and negative/blank wells Run replicates of sample controls where possible	Proper controls enable detection of technical variation and allow normalization across plates/runs.
5. Data Acquisition & QC	Monitor intra-run and inter-run coefficient of variation (CV) Check that the majority of proteins are within the panel's LLOQ-ULOQ (lower & upper limits of quantitation) Use control samples to detect drift or assay failure Use calibrator run in triplicate on each plate.	Ensures data quality and reliability; helps identify problematic samples or plates early.
6. Data Normalization & Analysis	Use absolute concentration units (pg/mL) when comparing groups or time-points; NPX units for relative changes if needed Apply normalization based on internal controls Handle values below LLOQ carefully (censor or impute appropriately) Use statistical methods suited to multiplex data (multivariate, multiple hypothesis correction)	Accurate comparisons, better interpretability, and avoidance of false positives/negatives.
7. Reporting & Interpreting Results	Include information on sample collection, handling, storage in Methods Report limits of detection / quantification for measured proteins Show CVs and QC metrics Interpret findings considering baseline levels in control mice, strain differences, age, sex	Transparency supports reproducibility; helps other labs understand context and limitations.

Practical Considerations & Common Pitfalls

Baseline vs induced levels: Understand what "normal" cytokine expression looks like in your mouse strain, age, sex. Panel document shows normal ranges for many proteins in control mice across multiple strains.

Inter-plate / Inter-operator variation: When multiple plates or operators are involved, use calibrators and controls to ensure comparability. The validation data from Olink show that inter-run CVs remain low when these are employed.

Assay saturation / hook effects: Very high protein concentrations can lead to decreased signal ("hook effect"). Make sure that expected cytokine levels fall within ULOQ. If not, dilution or alternate sampling points may be necessary.

Matrix effects & sample quality: Hemolysis, lipemia, and other interfering substances can skew readings (especially for low-abundance proteins). Where possible, avoid or test for such interference. ([Olink validation) data show that some proteins are sensitive to such interference.

Timeline & Cost-Planning

Sample prep & QC: depending on lab setup and sample number, allow time for sample collection, centrifugation, aliquoting, freezing. Likely ~1-2 days for moderate numbers.

Assay run: once samples and reagents are ready, the Olink protocol with the mouse panel has minimal hands-on time. Running on Signature Q100 reduces manual steps.

Data processing & analysis: QC and normalization typically take another 1-2 days depending on data volume and required statistical analysis. Include replication, outlier checking, etc.

Budget factors: reagent cost, instrument time, consumables, shipping (if using external service), data analysis support. Be sure to budget for QC samples and possibly custom validation if your sample matrix is unusual.

Conclusion & Call to Action

The Olink Target 48 Mouse Cytokine Panel offers a robust, high-precision tool for profiling immune mediators in preclinical research. With only 1 µL of sample, it allows measurement of ~43 well-validated cytokines and related immune modulators. This enables more comprehensive longitudinal studies, tighter experimental control, and better biological insight in non-clinical settings. If you are planning an immune response study, inflammation model, or translational project, integrating this panel in your workflow can reduce sample requirements, increase data richness, and improve reproducibility.

Next Steps

Request a custom quote from Creative Proteomics for using the Olink Target 48 Mouse Cytokine Panel in your project. We provide full support from sample handling to data delivery.

Consult with our experts to design your study (timepoints, sample types, controls) optimized for highest data quality.

Use our service offering for Olink proteomics analysis through Creative Proteomics; we also support integrated multi-omics studies if you wish to pair protein data with transcriptomics or metabolomics.

Thank you for reading. When you're ready, you can explore our related articles to deepen your understanding:

Let's collaborate to unlock new insights in your mouse model studies.

References

Monitoring Cerebrospinal Fluid Inflammatory Mediators by Olink Target 48 Cytokine Panel DOI: 10.1007/978-1-0716-4462-1_5
A Technical Evaluation of Plasma Proteomics Technologies — Beimers et al., 2025. DOI: https://doi.org/10.1021/acs.jproteome.5c00221
Correlations Within and Between Highly Multiplexed Proteomic Assays of Human Plasma — Rooney et al., Clin. Chem. 2025;71(6):677-687. https://doi.org/10.1093/clinchem/hvaf030
Noora Sissala, Haris Babačić, Isabelle R. Leo et al. Comparative evaluation of in-depth mass spectrometry and Olink Explore 3072 for plasma proteome profiling, 05 May 2025, PREPRINT (Version 1) available at Research Square . https://doi.org/10.21203/rs.3.rs-6501601/v1

* For research purposes only, not intended for clinical diagnosis, treatment, or individual health assessments.

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A Complete Guide to Olink Mouse Cytokine Panel