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NFκB p65 ELISA kit

 
ADI-EKS-446 2x96 wells 1,112.00 USD
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  • Fast- measure the active form of NFκB p65 in just 3 hours
  • Higher throughput - 96 well plate based method faster than EMSA and Western blot
  • Sensitive - chemiluminescent detection
The NFκB p65 EIA kit provides a sensitive, rapid and reliable method to measure the active form of NFκB p65. The assay uses streptavidin-coated plates with bound NFκB biotinylated-consensus sequence to capture only the active form of NFκB. The captured active NFκB is incubated with a specific NFκB p65 antibody, which is then detected using an HRP conjugated secondary antibody. The assay is developed with a chemiluminescent substrate and the signal is detected using a luminometer. Higher throughput 96 well plate based method faster than EMSA and Western blot with sensitive chemiluminescent detection.
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Product Specification

Assay Time:3 hours
 
Applications:ELISA, Chemiluminescence
 
Application Notes:For the quantitative determination of active NFκB p65 in cell lysates of human, mouse, and rat origin. Cited sample(s) include nuclear protein extracts.
 
Species reactivity:Human, Mouse, Rat
 
Use/Stability:Reagents require separate storage conditions upon receipt.
 
Shipping:Shipped on Dry Ice
 
Kit/Set Contains:Microtiter plate, Antibody, HRP Conjugate, Antibody dilution buffer, Wash buffer concentrate, Luminol enhancer solution, Stable peroxidase solution, Wild Type Competitor Duplex, Mutant Competitor Duplex, 5X Binding Buffer, 20X Poly dIdC, Actived HeLa Cell Nuclear Extract
 
Scientific Background:Stimulation of the canonical NFκB pathway leads to the phosphorylation of and subsequent degradation of IκB; phosphorylation of the latter is controlled by a specific set of proteins referred to as the I Kappa Kinases (IKK), which include IKKα and IKKβ. Under normal conditions, IκB acts as a key regulatory molecule in that it sequesters p65 or Rel A within the cytoplasm; this blocks NFκB dependent transcription. Upon phosphorylation of IκB by IKK’s, p65 is released and free to associate with the p50 subunit; p65 acts as a transactivator whereas p50 possesses DNA binding activity. Complex formation leads to the subsequent translocation of the NFκB complex into the nucleus and the concomitant up-regulation of NFκB dependent genes. Disruption of this signaling nexus has been documented in a number of human pathological conditions including inflammation, infection and cancer.
 
UniProt ID:Q04206 (human)
 

Product Literature References

Combined treatment of epigallocatechin gallate and Coenzyme Q10 attenuates cisplatin-induced nephrotoxicity via suppression of oxidative/nitrosative stress, inflammation and cellular damage: S. Fatima, et al.; Food Chem. Toxicol. 94, 213 (2016), Application(s): Determination of inflammation parameters, Rat NFkB levels were assessed, Abstract;
Identification of novel HIV-1 dependency factors in primary CCR4 + CCR6 + Th17 cells via a genome-wide transcriptional approach: A. Cleret-Buhot, et al.; Retrovirology 12, 102 (2015), Application(s): Specificity of NF-κB p65 DNA-binding was determined using wild-type and mutated NF-κB p65, Abstract; Full Text
Notch-1 signaling regulates astrocytic proliferation and activation after hypoxia exposure: Y. Zhang, et al.; Neurosci. Lett. 603, 12 (2015), Application(s): ELISA for NF-κB/p65 protein level analysis, Abstract;
Effects of curcumin on proinflammatory cytokines and tissue injury in the early and late phases of experimental acute pancreatitis: A. Gulcubuk, et al.; Pancreatology 13, 347 (2013), Application(s): Use of nuclear protein extracts, Abstract;
Vitamin A (retinol) downregulates the receptor for advanced glycation endproducts (RAGE) by oxidant-dependent activation of p38 MAPK and NF-kB in human lung cancer A549 cells: M.A. de Bittencourt Pasquali, et al.; Cell. Signal. 25, 939 (2013), Application(s): Use of nuclear protein extracts, Abstract;
Neuropilin-1 is expressed by breast cancer stem-like cells and is linked to NF-κB activation and tumor sphere formation: Y. Glinka, et al.; BBRC 425, 775 (2012), Application(s): Use of cell lysates, Abstract;
The curry spice curcumin selectively inhibits cancer cells growth in vitro and in preclinical model of glioblastoma: A. Zanotto-Filho, et al.; J. Nutr. Biochem. 23, 591 (2012), Application(s): Use of nuclear protein extracts, Abstract;
The preservative polyquaternium-1 increases cytoxicity and NF-kappaB linked inflammation in human corneal epithelial cells: T. Paimela, et al.; Mol. Vis. 18, 1189 (2012), Application(s): ELISA using cell lysates, Abstract; Full Text
NFκB inhibitors induce cell death in glioblastomas: A. Zanotto-Filho, et al.; Biochem. Pharmacol. 81, 412 (2011), Application(s): Use of nuclear protein extracts, Abstract;
Oestrogen treatment enhances the sensitivity of hormone-resistant breast cancer cells to doxorubicin: A.M. Scherbakov, et al.; Biosci. Rep. 31, 137 (2011), Application(s): Use of cell lysates, Abstract; Full Text
Capacity of omega-3 fatty acids or eicosapentaenoic acid to counteract weightlessness-induced bone loss by inhibiting NF-κB activation: From cells to bed rest to astronauts: S.R. Zwart, et al.; J. Bone Miner. Res. 25, 1049 (2010), Abstract;

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