Nf-kappa B Analysis
Nf-kappa B Analysis - Related Videos
14:58
Methods for the Modulation and Analysis of NF-κB-dependent Adult Neurogenesis
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Cited by 1 •
2014
Methods for the manipulation and analysis of NF-κB-dependent adult hippocampal neurogenesis are described. A detailed protocol is presented for a dentate gyrus-dependent behavioral test (termed the spatial pattern separation-Barnes maze) for the investigation of cognitive outcome in mice. This technique should also help enable investigations in other experimental settings.
02:26
NF-κB-dependent Signaling Pathway
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2021
The transcription factor NF-κB was discovered in 1986 in the lab of Nobel laureate Professor David Baltimore, for its interaction with the immunoglobulin light chain enhancer in B-cells. After more than three decades of study, it is now evident that NF-κB regulates the expression of over 100 genes. Most of these genes play an essential role in the innate and adaptive immune responses as well as the inflammatory responses of animals. NF-κB-dependent Signaling Mechanism The heterodimer of NF-κB...
08:51
Intraductal Injection of LPS as a Mouse Model of Mastitis: Signaling Visualized via an NF-κB Reporter Transgenic
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Cited by 14 •
2012
Described here is a technique in which lipopolysaccharide is injected into the lactating mouse mammary gland via the nipple to simulate mastitis, a condition commonly caused by bacterial infection. Lipopolysaccharide injection results in increased nuclear factor kappa B (NF-κB) signaling, visualized through bioluminescent imaging of an NF-κB luciferase reporter mouse.
10:57
NF-κB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells
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Cited by 4 •
2020
Here, we present a protocol to quickly and easily measure nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation in cell lines expressing NF-κB::luciferase reporter constructs, via measurements of luminescence in the cell lysate. Additionally, gene expression is determined via RT-qPCR isolated from cells infected with Salmonella Typhimurium.
15:04
A Neuronal and Astrocyte Co-Culture Assay for High Content Analysis of Neurotoxicity
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Cited by 46 •
2009
This article describes a novel protocol and reagent set designed for sensitive measurement of neurotoxic effects of compounds and treatments on co-cultures of neurons and astrocytes using high content analysis. Results demonstrate that high content analysis represents an exciting novel technology for neurotoxicity assessment.
FAQs
The initiating signal provides essential context for the observed response. Pattern-recognition receptors, cytokine receptors, and cellular stress can all engage the pathway, but they represent different biological inputs. Identifying the stimulus helps investigators relate NF-kappa B activity to host sensing, inflammatory signaling, or stress responses and interpret how that input may influence immune gene expression.
Nuclear localization indicates that NF-kappa B has moved from the cytoplasm toward the site of DNA regulation, whereas target-gene expression reflects a downstream transcriptional outcome. These measurements therefore examine different points in the response. Assessing both can help connect pathway activity with the resulting immune or inflammatory gene program rather than relying on one readout alone.
The analysis can separate early pathway events from later biological consequences by examining distinct readouts. Changes in IκB regulation and NF-kappa B nuclear localization reflect pathway control, DNA binding indicates interaction with target sequences, and target-gene expression reflects downstream effects. This organization helps clarify whether a response is altered at activation, DNA regulation, or transcriptional output.
A practical workflow begins by defining the relevant cellular stimulus, then examining a selected NF-kappa B readout in resting and stimulated conditions. Researchers may assess nuclear localization, DNA binding, or expression of NF-kappa B target genes. Comparing these measurements with the experimental condition helps reveal how the stimulus changes pathway activity and immune-related transcription.
This analysis is useful when researchers need to determine how pathogens or host-derived signals shape immune and inflammatory responses. Measurements can connect an experimental condition with NF-kappa B localization, DNA binding, or target-gene expression. The resulting information supports investigation of inflammation, disease mechanisms, and possible therapeutic interventions involving pathway regulation.
By measuring NF-kappa B responses after exposure to relevant pathogen-associated or host signals, investigators can examine how those conditions influence immune gene regulation. Nuclear localization, DNA binding, and target-gene expression provide complementary evidence about pathway behavior. These results can help relate altered NF-kappa B activity to inflammatory responses and broader disease mechanisms in infection research.