SigPath™ NFκB Phosphorylation Assay Kit

CAT#: ITS-1125-WB21
Product Type: Assay Kit
Species: Human, mouse
Target: Phosphorylated NFκB (p65) subunit at the serine 536 residue.
Brand: SigPath
Development Stage: Research-Grade Signal Pathway Assay Development
Short Description
Quantifies NFκB (p65) phosphorylation at Ser536 in cell lysates using an ELISA-based colorimetric method.
Brand
SigPath
Development Stage
Research-Grade Signal Pathway Assay Development
Description
The SigPath™ NFκB Phosphorylation Assay Kit provides a streamlined, cell-based ELISA for quantifying p65 (Ser-536) phosphorylation in cultured cells. By eliminating cell lysis and incorporating total protein normalization, this high-throughput system ensures precise measurement of NF-κB activation. It is an essential tool for screening anti-inflammatory drug candidates and studying complex signaling kinetics in pharmaceutical research.
Features
High Sensitivity & Specificity: Employs advanced ELISA-based sandwich technology to detect low-abundance phosphorylated NFκB p65 with exceptional signal-to-noise ratios.
Streamlined Homogeneous Format: Features a rapid, simplified workflow that minimizes manual handling, ensuring high reproducibility and scalability for automation.
Universal Sample Compatibility: Optimized for diverse biological matrices, including cell lysates and tissue homogenates, facilitating broad translational research applications.
Quantitative Signal Detection: Delivers precise colorimetric or fluorometric data output, allowing for rigorous kinetic modeling and comparative statistical analysis.
Process Relevance
Quantifying p65 phosphorylation to evaluate inflammatory signaling modulation and assess the potency of NFκB pathway inhibitors.
Application Stage
Target validation, high-throughput lead optimization, and pre-clinical efficacy monitoring in drug discovery pipelines.
Applications
Assessing nuclear translocation and activation across multiple cell lines, screening small molecule libraries, and investigating cytokine-induced inflammatory responses in complex primary cell cultures.
Qualified With
Internal performance validation using reference standards under defined assay conditions.
Target
Phosphorylated NFκB (p65) subunit at the serine 536 residue.
Detection Method
Fluorescence Detection (Ex 530 nm / Em 585 nm or Ex 360 nm / Em 452 nm)
Sample Type
Cells
Research Areas
Oncology, immunology, chronic inflammatory diseases, drug discovery, cardiovascular pathology, etc.
Size
100 tests
Data Output Type
Quantitative protein phosphorylation measurement via absorbance or fluorescence intensity.
Assay Time
6.5 h
Species
Human, mouse
Storage
Store at −20 °C; upon receipt to maintain reagent stability. Follow recommended handling procedures to ensure consistent assay performance.
Storage Comment
Avoid repeated freeze-thaw cycles. During use, minimize the time components are exposed to room temperature and return them promptly to −20 ℃ storage after handling. Do not store at elevated temperatures for extended periods.
Expiry Date
6 months under recommended storage conditions to ensure optimal performance.
Note
For bulk pricing or custom reagent inquiries, please contact us by email or phone. Products are shipped on ice via FedEx.
Restrictions
For Research Use Only (RUO). Not intended for diagnostic or therapeutic use.
Shipping
Shipped on ice under temperature-controlled conditions.
Background
The NFκB complex acts as a central transcription factor governing immune homeostasis and cell survival. Dysregulation of its phosphorylation status drives oncogenesis and autoimmune pathologies, making it a critical biomarker.
Pathways
Activation involves IκB kinase mediated phosphorylation, triggering IκB degradation and p65 nuclear translocation. This pathway regulates downstream pro-inflammatory cytokines, anti-apoptotic factors, and secondary immune response mediators.
For Research Use Only | Not For Clinical Use
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