NF-κB Signaling


NF-kappaB Overview

Nuclear Factor kappa-light-chain-enhancer of activated B cells or NF-κB is found in almost all animal cells and is a protein complex that controls the transcription of DNA. This protein complex is involved in numerous cellular responses and plays a key role in regulating the immune response to infection.

NF-κB is also implicated in processes of synaptic plasticity and memory that require additional avenues of research to flesh out completely the possibilities and uses of this compound.








NF-κB button


NF-κB Structure

All proteins of the NF-κB family share a Rel homology domain in their N-terminus. A subfamily of NF-κB proteins, including RelA, RelB, and c-Rel, have a transactivation domain in their C-termini. In contrast, the NF-κB1 and NF-κB2 proteins are synthesized as large precursors, undergoing processing to generate mature and active NF-κB subunits.

NF-κB is detected in most cell types, and specific NF-κB binding sites are identified in promoters and enhancers of increasing number of inducible genes. The transcription factor NF-κB consists of homo- or heterodimers of different subunits, members of a family of structurally related proteins – Rel / NF-κB proteins. Rel proteins contain a conserved N-terminal region, called the Rel Homology Domain (RHD), which contains the DNA-binding and dimerization domains and the nuclear localization signal of the Rel proteins.



NF-κB Activation

NF-κB can be activated by a number of stimuli, including exposure of cells to Lipopolysaccharides or inflammatory cytokines such as Tumor Necrosis Factor or Interleukin-1, growth factors, lymphokines, oxidant-free radicals, inhaled particles, viral infection or expression of certain viral or bacterial gene products.  Activation of NF-κB occurs according to specific and sequential phosphorylation of residues such as Ser32 and Ser36 of IκBa leading to proteasome - mediated degradation.  Further, phosphorylation of Ser 276 on RelA/p65 allows its oligomerization and reulation of DNA-binding.  NF-κB phosphorylation is crucial for the regulation of the NF-κB Activation.



Significance and Uses of NF-κB

NF-κB is important in regulating cellular responses because it belongs to the category of "rapid-acting" primary transcription factors. These transcription factors are present in cells in an inactive state and do not require new protein synthesis to be activated, allowing NF-κB to be a first and fast responder to harmful cellular stimuli. Receptor activator of nuclear factor kappa B, which is a type of TNFR, is a central activator of NF-κB. Many bacterial products and stimulation of a wide variety of cell-surface receptors lead to NF-κB activation and fairly rapid changes in gene expression. 



NF-κB and Cancer Research

NF-κB is widely used by eukaryotic cells as a regulator of genes that control cell proliferation and cell survival, thus many human tumors have unregulated or non-regulated NF-κB. NF-κB is extremely useful to identify and to understand how cancer is regulated. Active NF-κB turns on the expression of genes that keep the cell proliferating and protect the cell from conditions that would otherwise cause its death through apoptosis mechanism.

In tumor cells, NF-κB is active either due to mutations in genes encoding the NF-κB transcription factors themselves or in genes that control NF-κB activity. Blocking NF-κB can cause tumor cells to stop proliferating, to die, or to become more sensitive to the action of anti-tumor agents. Therefore, NF-κB is the subject of much active research among pharmaceutical companies as a target for anti-cancer therapy.



Other NF-κB Research Areas

NF-κB controls many genes involved in inflammation and is found to be chronically active in many inflammatory diseases, such as inflammatory bowel disease, arthritis, sepsis, gastritis, asthma, atherosclerosis. Many natural products described as having anti-cancer and anti-inflammatory activity have been shown to inhibit NF-κB. Recent has highlighted the importance of the connection between NF-κB, inflammation, and cancer, while underscoring the value of therapies that regulate the NF-κB activity.



   TrueBlot IP/Western Blot Mouse Rabbit Goat

 

 

 

 

 

   

 

 



NF-κB (p50) Transcription Factor Assay Kit


NFKB p50 ELISA  

 

Rockland’s NF-κB (p65) Transcription Factor Assay is a non-radioactive, sensitive method for detecting specific transcription factor DNA binding activity in nuclear extracts and whole cell lysates. A 96-well enzyme-linked immunosorbent assay (ELISA) replaces the cumbersome radioactive electrophoretic mobility shift assay (EMSA). A specific double stranded DNA (dsDNA) sequence containing the NF-κB response element is immobilized to the wells of a 96 well plate. NF-κB contained in a nuclear extract, binds specifically to the NF-κB response element. NF-κB (p65) is detected by addition of specific primary antibody directed against NF-κB (p65). A secondary antibody conjugated to HRP is added to provide a sensitive colorimetric readout at 450 nm. Rockland’s NF-κB (p65) Transcription Factor Assay detects human NF-κB (p65). It will not cross-react with NF-κB (p50).




Phospho Related Antibodies