Signal Transduction Assay
Trace Compound Activity Through the Signaling Pathway
A signal transduction assay measures how a compound's activity at its target translates into downstream biological effect, tracing pathway activation or inhibition from early phosphorylation events and second-messenger signaling through transcriptional and phenotypic output. WuXi Biology and Testing in vitro platform covers various pathways, such as Ras/Raf/MEK/ERK, JAK/STAT, and DNA damage response signaling, using validated readouts and multiplex detection technologies such as RT-qPCR, Western blot, in-cell Western, TR-FRET (HTRF), and AlphaLISA.
Pre-built Ready-to-Go assays and disease-relevant mutant panels allow projects to move rapidly from hit identification to detailed mechanism-of-action characterization. These assays pair directly with our Target Engagement Assays to confirm that compound binding produces the intended functional and phenotypic outcome.
Cell Signaling Pathway Assays
Confirm that a compound's target engagement translates into the intended downstream biological effect. Our signal transduction platform profiles pathway activation and inhibition across various pathways, such as:
- Ras/Raf/MEK/ERK pathway quantifies pathway activation and inhibitor potency across RAS mutant panels (G12C, G12D, G12V, and more) using p-ERK and viability readouts
- JAK/STAT pathway profiles cytokine-driven phosphorylation across STAT1–STAT6 isoforms in response to IL-2, IL-4, IL-6, IL-12, and IFN-γ, among other immunological stimuli
- DNA damage response (DDR) measures PARP trapping, PARylation, and γH2AX/p-H2AX induction, EdU Incorporation, senescence assays to characterize DDR inhibitor mechanism and selectivity
- Nuclear receptor and stress pathway signaling assesses GPCR, Wnt/β-catenin, p53, hypoxia, and endoplasmic reticulum stress pathway activity using validated reporter cell lines
- Covalent compound analysis determines kinact/KI kinetics, off-target binding sites, and proteome-wide occupancy for irreversible inhibitors
Signal Transduction Detection Technologies
We match the detection platform to the pathway and sensitivity required, drawing on a validated set of reporter, molecular, and proximity-based technologies, including everything in the following table.
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Frequently Asked Questions
A signal transduction assay measures how a compound's activity at its target translates into downstream biological effect, tracking pathway activation or inhibition through phosphorylation events, transcriptional output, or other functional endpoints inside the cell.
Yes. Signal transduction assays can quantify both cascade activation and target inhibition by monitoring changes in phosphorylation, second-messenger levels, cytokine secretion, transcriptional regulation, or other downstream functional cellular responses.
We support major oncology and immunology pathway families, including Ras/Raf/MEK/ERK, JAK/STAT (STAT1–STAT6), DNA damage response, GPCR signaling, Wnt/β-catenin, p53, hypoxia, and endoplasmic reticulum stress signaling, using validated reporter cell lines and pathway-specific antibodies.
Depending on the pathway and sensitivity required, we use luciferase-based gene reporter assays, RT-qPCR, Western blot and in-cell Western, TR-FRET (HTRF), ELISA, AlphaLISA, and multiplex cytokine detection platforms such as MSD,and Luminex.
Yes. We maintain validated cell line panels covering common oncogenic drivers — including KRAS (G12C, G12D, G12V, and diverse alleles), EGFR mutations, FGFR mutations, and BRAF V600E, etc. so pathway modulation can be measured directly in a disease-relevant genetic background.
Yes. Pathway-level readouts are frequently generated alongside our Target Engagement Assays to confirm that a compound's binding translates into the intended functional and phenotypic outcome.
Yes. We offer PARP trapping, PARylation, and γH2AX/p-H2AX, EdU Incorporation, senescence assays to characterize DDR inhibitor mechanism, potency, and selectivity, including in genetically defined MSI/MSS cell line panels.