NF-κB Signaling Pathway

Nuclear factor-κB (NF-κB) belongs to a family of transcription factors that play pivotal roles in inflammatory responses and immunological reactions. NFκB pathway can be activated by a variety of stimuli, including TNF-α (tumor necrosis factor α), interleukin 1 (IL-1), T and B cell mitogens, bacterial lipopolysaccharide (LPS) and viral proteins. Eurofins DiscoverX offers a simple one-step IκB degradation and NFκB reporter and translocation assay to study the NFκB pathway. These fully optimized, target-validated cell lines provide you convenience, relevant hits and guaranteed performance for screening, profiling or research applications.

 

Features and Benefits
 

  • Short compound incubation times (30-60 minutes); reduce toxicity & false positives

  • Direct, non-reporter gene, functional Assay

  • Monitor degradation of IκB

  • Whole cell, HTS friendly chemiluminescent assay

  • Transcription independent assay

  • Increased specificity

  • Available in multiple backgrounds


Applications
 

  • Screen inhibitors for: TNFα signaling, IκB  kinases

Assay List

Cell Lines

Target Description Cat. No.
RANK-NFκB PathHunter® U2OS RANK-NF-KB Reporter Cell Line 93-1156C3
NFκB PathHunter® U2OS NF-KB Reporter Cell Line 93-1157C3
IκB PathHunter® HEK 293 IκB Degradation Cell Line 93-0538C1
IκB PathHunter® THP-1 IκB Degradation Cell Line 93-0538C14
IκB PathHunter® A549 IκB Degradation Cell Line 93-0538C15
IκB PathHunter® U2OS IκB Degradation Cell Line 93-0538C3
IκB PathHunter® Jurkat IκB Degradation Cell Line 93-1121C19
RANK-IκB PathHunter® U2OS RANK-IκB Functional Assay 93-0994C3
RELA PathHunter® HEK 293 RELA Nuclear Translocation Cell Line 93-0907C1

Assay Ready Kits
Target Description Cat. No.
IκB PathHunter® Adalimumab Bioassay Kit 93-0538B15-00131
IκB PathHunter® Adalimumab Bioassay Kit 93-0538B15-00132
IκB PathHunter® eXpress HEK 293 IκB Degradation Assay 93-0538E1CP7L
IκB PathHunter® eXpress HEK 293 IκB Degradation Assay 93-0538E1CP7M
IκB PathHunter® eXpress HEK 293 IκB Degradation Assay 93-0538E1CP7S
RELA PathHunter® eXpress HEK 293 RELA Nuclear Translocation Assay 93-0907E1CP0L
RELA PathHunter® eXpress HEK 293 RELA Nuclear Translocation Assay 93-0907E1CP0M
RELA PathHunter® eXpress HEK 293 RELA Nuclear Translocation Assay 93-0907E1CP0S

 

Assay Principle

NF-κB Signaling Pathway Reporter assay principle showing NF-κB signaling reporter construct regulates PathHunter EFC reporter expression via upstream NF-κB response elements controlled by NF-κB transcription factor binding. Upstream signaling events result in NF-κB nuclear translocation and subsequent response-element binding. Signaling activity is measured by monitoring PathHunter EFC Reporter abundance after complementary enzyme acceptor (EA) addition, resulting in enzyme fragment complementation to create an active β-gal enzyme, capable of substrate hydrolysis and production of a chemiluminescent signal.

How it Works


PathHunter® U2OS NF-κB Reporter Assay




Eurofins DiscoverX offers PathHunter® IkB functional assays in which IkB (Inhibitor of κB)is tagged with small ProLabel peptide. The assay measures human IκB degradation in response to activation of the NFkB pathway. In the presence of TNFα or other compounds, cells signal through downstream  IκB proteins. IκB protein is degraded in response to compound stimulation.

How it Works