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Our scientists can help you choose the right assay format, substrate, and enzyme for your experiment.
About MAPK14
MAPK14, also known as p38α, is a member of the mitogen-activated protein kinase (MAPK) family, a group of serine/threonine kinases that transduce extracellular stress signals into coordinated cellular responses. MAPK14 is activated through a canonical three-tier kinase cascade: upstream MAP3Ks phosphorylate MKK3 and MKK6, which in turn dual-phosphorylate MAPK14 on the Thr180-Tyr182 activation loop motif. Key substrates include MK2 (MAPKAPK2), ATF2, MSK1, and HSP27, linking MAPK14 to inflammation, cell cycle arrest, apoptosis, and cytokine production. The kinase is a central node in innate immune signaling downstream of Toll-like receptors, TNF-α, and IL-1β. Dysregulation of MAPK14 is implicated in rheumatoid arthritis, inflammatory bowel disease, cardiovascular disease, neurodegeneration, and multiple cancers. Its well-validated role in driving pro-inflammatory cytokine synthesis (particularly TNF-α and IL-6) makes MAPK14 an attractive therapeutic target, and it has been extensively pursued by the pharmaceutical industry for both acute and chronic inflammatory indications.
Why PhosphoSens for MAPK14?
MAPK14 drug discovery presents several assay challenges: the kinase is subject to feedback regulation and requires careful activation-state validation; physiological ATP concentrations (1–5 mM) are critical because many p38α inhibitor scaffolds are ATP-competitive and IC50 values shift dramatically at non-physiological ATP. Traditional endpoint methods such as ADP-Glo and HTRF capture single time points, missing biphasic kinetics or compound-induced substrate aggregation. Radiometric assays offer sensitivity but lack throughput and real-time resolution. AssayQuant's PhosphoSens continuous fluorescent assay monitors MK2-derived peptide substrate phosphorylation in real time at physiological ATP, generating full progress curves. This enables direct measurement of kinact/KI for covalent inhibitors, accurate Km/Vmax determination, and early detection of assay artifacts — all without antibodies — dramatically improving data quality and throughput for MAPK14 inhibitor profiling.
Key Pathways:
p38 MAPK Signaling
Toll-Like Receptor Signaling
TNF-alpha/NF-kB Signaling
Stress-Activated Protein Kinase Cascade
Talk to a Scientist
Have questions about MAPK14 assay design, selectivity panels, or covalent inhibitor characterization?
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How pricing works:
Select a kit or substrate below, then request a quote. Bundle discounts apply automatically.
Select a kit or substrate below, then request a quote. Bundle discounts apply automatically.
Real-Time MAPK14 Activity Assays
Continuous, real-time fluorescent assays optimized for quantitative MAPK14 activity measurements, IC50 determination, and mechanistic studies.
Assay Format: Continuous kinetic fluorescence (real-time)
PhosphoSens-Kinetic assays directly quantify enzyme activity by continuously monitoring substrate phosphorylation or dephosphorylation in real time, generating a full progress curve in every well.
Learn more about PhosphoSens-Kinetic →Need pricing or availability? Select a kit or substrate to request a quote below.
Kits
PhosphoSens-Kinetic Kinase Activity Kit - AQT0490
Ready-to-use assay kits containing substrate and all essential reagents.
Kit Contents
- Optimized PhosphoSens® substrate (AQT0490)
- ATP, DTT, EGTA, Reaction Buffer & Dilution Buffer
Automatically save 10% when bundling 10ug recombinant enzyme with your 1,000 assay kit. View enzymes
Price: $425
Substrate
PhosphoSens Kinase Substrate AQT0490
Bulk PhosphoSens® substrate for assay development and high-throughput workflows.
Specifications
Automatically save 10% when bundling Reagent Packs with your substrate. View Reagent Packs
Price: $2795
Ready to request pricing for MAPK14?
Select a kit, substrate, or enzyme above. Our team will confirm pricing, availability, and any applicable bundle discounts.
Request a QuoteTRF MAPK14 Screening Assays
Robust endpoint TRF assays optimized for high-throughput screening and quantitative MAPK14 activity measurement in plate-based workflows.
Assay Format: Time-resolved fluorescence (TRF, endpoint)
PhosphoSens-Red assays use europium-based time-resolved fluorescence detection to enable robust, plate-based quantification of enzyme activity, making them well suited for high-throughput screening workflows.
Learn more about PhosphoSens-Red →Need pricing or availability? Select a kit or substrate to request a quote below.
Kits
PhosphoSens-Red Kinase Activity Kit - AQT0649
Ready-to-use TRF assay kits containing substrate and essential reagents.
Kit Contents
- Optimized PhosphoSens® substrate (AQT0649)
- ATP, DTT, EGTA, Reaction Buffer & Dilution Buffer
- Europium Detection Solution
Automatically save 10% when bundling 10ug recombinant enzyme with your 1,000 assay kit. View enzymes
Price: $455
Substrate
PhosphoSens Kinase Substrate AQT0649
Bulk substrate for PhosphoSens-Red TRF assays and high-throughput workflows.
Specifications
Automatically save 10% when bundling Reagent Packs with your substrate. View Reagent Packs
Price: $2795
Ready to request pricing for MAPK14?
Select a kit, substrate, or enzyme above. Our team will confirm pricing, availability, and any applicable bundle discounts.
Request a QuoteNo recombinant enzymes are currently available for MAPK14.
Frequently Asked Questions: MAPK14 Assays
Why is physiological ATP concentration important when profiling MAPK14 inhibitors?
Most MAPK14 inhibitors are ATP-competitive, meaning their measured IC50 values are highly sensitive to the ATP concentration used in the assay. Assays run at sub-physiological ATP (e.g., Km ATP) can underestimate IC50 by 10-fold or more compared to cellular conditions. AssayQuant's PhosphoSens platform supports assays at physiological ATP (1u20135 mM), providing inhibitor potency data that better translates to cell-based and in vivo settings.
Can PhosphoSens detect allosteric and type II inhibitors of MAPK14 such as BIRB 796?
Yes. Because PhosphoSens monitors phosphorylation continuously in real time, it captures the slow-binding kinetics characteristic of type II inhibitors like BIRB 796 (Doramapimod) that are missed by endpoint assays taken at a fixed time. Full progress curves reveal residence time effects and slow kon/koff behavior, enabling more accurate mechanistic classification of MAPK14 inhibitor binding modes.
What substrate does AssayQuant recommend for MAPK14 activity assays?
AssayQuant offers PhosphoSens peptide substrates derived from validated MAPK14 substrates, including sequences based on the MK2 and ATF2 phosphorylation sites, which are efficiently recognized by activated MAPK14. These substrates incorporate the proprietary Sox-based fluorophore that generates a real-time fluorescent signal upon phosphorylation, enabling continuous kinetic monitoring without antibodies or secondary detection reagents.
Data & Resources
Explore data and documents to support your kinase and phosphatase experiments. Download sample data, protocols and other resources to see how our assays perform and to help you get started in your own lab. All validation data generated using PhosphoSens® assays under recommended conditions.
Sample Report: AQT0734 - EGFR Assay Validation Report
Each validation report provides experimental conditions and data showing:
- Enzyme titration and linearity
- Sensor peptide substrate titration and Km determination
- ATP titration and Km determination
- DMSO titration and assay tolerance
- Inhibitor titration and IC50 determination
Protocol
Inhibitor IC50 Protocol
See how the PhosphoSens-Kinetic Assay can be used to find the IC50 of a kinase inhibitor.
Download Protocol →Webinar
Webinar: Continuous Assay Validation
Discover how continuous assay formats power deep understanding of kinase function. See how PhosphoSens® assays guide inhibitor profiling, selectivity assessment, and mechanistic characterization.
Watch the Webinar →Service
KinSight™ MAPK14 Profiling
Need broader selectivity data? KinSight profiling runs your compounds across our full kinase panel under identical PhosphoSens conditions.
Learn about KinSight →Stay Informed
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