AB210613

NFkB p65 Transcription Factor Assay Kit (Colorimetric)

Name: NFkB p65 Transcription Factor Assay Kit (Colorimetric) (ab210613) | Abcam
Brand: Abcam Limited
SKU: AB210613
Price: 1435 USD
Availability: InStock

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(9 Publications)

NFkB p65 Transcription Factor Assay Kit (Colorimetric) (ab210613) is a high throughput assay to quantify NFkB p65 activation in nuclear extracts.

View Alternative Names

NFKB3, RELA, Transcription factor p65, Nuclear factor NF-kappa-B p65 subunit, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3

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Functional Studies - NFkB p65 Transcription Factor Assay Kit (Colorimetric) (AB210613)

FuncS

Supplier Data

Functional Studies - NFkB p65 Transcription Factor Assay Kit (Colorimetric) (AB210613)

Unstimulated Jurkat (Gray) and stimulated Jurkat (TPA + CI) (Black) cells were tested for NFkB p65 activation.

Different amounts of nuclear extracts from unstimulated Jurkat (white) and stimulated with TPA and calcium ionophore (black) were tested for NFkB p65 activation. This data is provided for demonstration only.

FuncS

Supplier Data

Functional Studies - NFkB p65 Transcription Factor Assay Kit (Colorimetric) (AB210613)

Nuclear extracts (10 μg/well) prepared from a variety of cell lines were tested for NFkB p65 activity. Cell lines were untreated HeLa cells, HeLa cells treated with TNFα, untreated Jurkat cells, Jurkat cells treated with PMA and calcium ionophore (CI), and Raji cells. These results are provided for demonstration only.

Key facts

Detection method

Colorimetric

Sample types

Nuclear Extracts

Reacts with

Rat, Human, Mouse

Results type

Semi-Quantitative

Sensitivity

< 500 ng/well

Assay time

3h 30m

Assay Platform

Microplate reader

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Product details

NFkB p65 Transcription Factor Assay Kit (Colorimetric) (ab210613) is a high throughput assay to quantify NFkB p65 activation in nuclear extracts. This assay combines a quick ELISA format with a sensitive and specific non-radioactive assay for transcription factor activation.

A specific double stranded DNA sequence containing the NFkB p65 consensus binding site (5' – GGGACTTTCC – 3') has been immobilized onto a 96-well plate. Active NFkB p65 present in the nuclear extract specifically binds to the oligonucleotide. NFkB p65 is detected by a primary antibody that recognizes an epitope of NFkB p65 accessible only when the protein is activated and bound to its target DNA. An HRP-conjugated secondary antibody provides sensitive colorimetric readout that at OD 450 nm. This product detects human, mouse and rat NFkB p65.

Key performance and benefits:

Assay time: 3.5 hours (cell extracts preparation not included).
Detection limit: < 0.5 μg nuclear extract/well.
Detection range: 0.2 – 10 μg nuclear or whole cell extract/well.

The transcription factor NFkB is implicated in the regulation of many genes that code for mediators of the immune, acute phase and inflammatory responses. The DNA-binding protein complex recognizes a discrete nucleotide sequence (5´-GGGACTTTCC-3´) in the upstream region of a variety of cellular and viral response genes. NFkB is composed of homo- and heterodimeric complexes of members of the Rel (NFkB) family. There are five subunits of the NFkB family in mammals: p50, p65 (RelA), c-Rel, p52 and RelB. These proteins share a conserved 300 amino acid sequence in the N-terminal region, known as the Rel homology domain, that mediates DNA binding, protein dimerization and nuclear localization. This domain is also a target of the IkB inhibitors, which include IkBα, IkBβ, IkBγ, Bcl-3, p105 and p100. Various dimer combinations of the NFkB subunits have distinct DNA binding specificities and may serve to activate specific sets of genes such as adhesion molecules, immunoreceptors and cytokines. The p50/p65 (NFkB1/RelA) heterodimers and the p50 homodimers are the most common dimers found in the NFkB signaling pathway. In the majority of cells, NFkB exists in an inactive form in the cytoplasm, bound to the inhibitory IkB proteins. Treatment of cells with various inducers results in the phosphorylation, ubiquitination and subsequent degradation of IkB proteins. Proteolytic cleavage of p105 results in two proteins: p50, which has DNA-binding activity but no transactivation domain, and its antagonist, the inhibitory IkBg protein. This results in the release of NFkB dimers, which subsequently translocate to the nucleus, where they activate appropriate target genes. NFkB can be activated by a number of stimuli, including components of bacterial cell walls, such as lipopolysaccharide, or inflammatory cytokines, such as TNF-α or IL-1β.

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What's included?

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Properties and storage information

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

Multi

Appropriate long-term storage conditions

Multi

Storage information

Please refer to protocols

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

NF-kB p65 also known as RelA is a significant component of the NF-kB protein complex. This complex usually involves a molecular weight for p65 of approximately 65 kDa. NF-kB p65 is mechanistically a transcription factor that regulates genes involved in inflammation cell survival and immune response. Expression of p65 is widely seen in various cell types including immune cells and epithelial cells suggesting its role in numerous physiological processes.

Biological function summary

NF-kB p65 acts as part of the larger NF-kB complex usually forming a heterodimer with other family members like p50. This complex translocates to the nucleus upon activation where it binds specific DNA sequences to regulate gene expression. The p65 subunit is essential for transactivating target genes involved in immune and inflammatory responses. Its regulation is important for proper cellular functioning especially in the maintenance of immune homeostasis and inflammatory response.

Pathways

NF-kB p65 participates in critical pathways like the NF-kB signaling and Toll-like receptor pathways. These pathways are fundamental for initiating immune responses and contribute to the regulation of apoptosis and cellular stress responses. In the context of these pathways the IKK complex plays a pivotal role in NF-kB activation leading to the phosphorylation and subsequent degradation of inhibitors that retain NF-kB p65 in the cytoplasm therefore allowing its movement to the nucleus.

NF-kB p65 is implicated in conditions such as cancer and autoimmune diseases. Its dysregulation can lead to persistent activation of inflammatory pathways contributing to tumorigenesis and chronic inflammation. In the context of these diseases proteins such as the p50 subunit and the IKK complex are often involved reflecting the importance of tight regulation of NF-kB signaling in preventing pathological conditions.

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Product protocols

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Target data

NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52. The heterodimeric RELA-NFKB1 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. The NF-kappa-B heterodimeric RELA-NFKB1 and RELA-REL complexes, for instance, function as transcriptional activators. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. The inhibitory effect of I-kappa-B on NF-kappa-B through retention in the cytoplasm is exerted primarily through the interaction with RELA. RELA shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Besides its activity as a direct transcriptional activator, it is also able to modulate promoters accessibility to transcription factors and thereby indirectly regulate gene expression. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. Essential for cytokine gene expression in T-cells (PubMed : 15790681). The NF-kappa-B homodimeric RELA-RELA complex appears to be involved in invasin-mediated activation of IL-8 expression. Key transcription factor regulating the IFN response during SARS-CoV-2 infection (PubMed : 33440148).

See full target information RELA

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Publications (9)

Recent publications for all applications. Explore the full list and refine your search

Frontiers in pharmacology 16:1546010 PubMed40606616

2025

Innovative pH-responsive alginate-coated rosuvastatin-loaded chitosan nanoparticles: a targeted approach to inhibiting HMGB1-activated RAGE/TLR4-NFκB signaling in colonic inflammation in rats.

Applications

Unspecified application

Species

Unspecified reactive species

Elsayed A Elmorsy,Sameh Saber,Zubida M Al-Majdoub,Rabab S Hamad,Mustafa Ahmed Abdel-Reheim,Asmaa Ramadan,Norah Suliman Alsoqih,Mariam S Alharbi,Hamad Alsaykhan,Alshaimaa A Farrag,Hanan Eissa,Rasha Abdelhady,Abousree T Ellethy,Mostafa M Khodeir,Hossam A Elsisi,Syed Suhail Ahmed,Ahmed Y Kira

Science advances 11:eadr2226 PubMed40315317

2025

A neuroimmune pathway drives bacterial infection.

Applications

Unspecified application

Species

Unspecified reactive species

Nian Wang,Jiao Liu,Runliu Wu,Feng Chen,Ruoxi Zhang,Chunhua Yu,Herbert Zeh,Xianzhong Xiao,Haichao Wang,Timothy R Billiar,Ling Zeng,Jianxin Jiang,Daolin Tang,Rui Kang

BMC medicine 21:287 PubMed37542259

2023

Decreased TMIGD1 aggravates colitis and intestinal barrier dysfunction via the BANF1-NF-κB pathway in Crohn's disease.

Applications

Unspecified application

Species

Unspecified reactive species

Longyuan Zhou,Liguo Zhu,Xiaomin Wu,Shixian Hu,Shenghong Zhang,Min Ning,Jun Yu,Minhu Chen

British journal of pharmacology 180:308-329 PubMed36166825

2022

Icariside II preconditioning evokes robust neuroprotection against ischaemic stroke, by targeting Nrf2 and the OXPHOS/NF-κB/ferroptosis pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Jianmei Gao,Congjian Ma,Dianya Xia,Nana Chen,Jianyong Zhang,Fan Xu,Fei Li,Yuqi He,Qihai Gong

FASEB journal : official publication of the Federa 33:10393-10408 PubMed31233346

2019

Isorhynchophylline exerts antidepressant-like effects in mice modulating neuroinflammation and neurotrophins: involvement of the PI3K/Akt/GSK-3β signaling pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Yan-Fang Xian,Siu-Po Ip,Hui-Qin Li,Chang Qu,Zi-Ren Su,Jian-Nan Chen,Zhi-Xiu Lin

Journal of thermal biology 83:69-79 PubMed31331527

2019

Extract from Coriolus versicolor fungus partially prevents endotoxin tolerance development by maintaining febrile response and increasing IL-6 generation.

Applications

Unspecified application

Species

Unspecified reactive species

Tomasz Jędrzejewski,Jakub Piotrowski,Małgorzata Pawlikowska,Sylwia Wrotek,Wieslaw Kozak

Cells, tissues, organs 206:62-72 PubMed30716735

2019

Ameliorative Effect of Cardamom Aqueous Extract on Doxorubicin-Induced Cardiotoxicity in Rats.

Applications

Unspecified application

Species

Unspecified reactive species

Maha Abu Gazia,Mohammed Abu El-Magd

Diabetology & metabolic syndrome 10:89 PubMed30534206

2018

Antidiabetic efficacy of lactoferrin in type 2 diabetic pediatrics; controlling impact on PPAR-γ, SIRT-1, and TLR4 downstream signaling pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Waleed A Mohamed,Mona F Schaalan

The Journal of biological chemistry 293:10561-10573 PubMed29802199

2018

C6orf106 is a novel inhibitor of the interferon-regulatory factor 3-dependent innate antiviral response.

Applications

Unspecified application

Species

Unspecified reactive species

Rebecca L Ambrose,Yu Chih Liu,Timothy E Adams,Andrew G D Bean,Cameron R Stewart

View all publications

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NFkB p65 Transcription Factor Assay Kit (Colorimetric)

Functional Studies - NFkB p65 Transcription Factor Assay Kit (Colorimetric) (AB210613)

Functional Studies - NFkB p65 Transcription Factor Assay Kit (Colorimetric) (AB210613)

Key facts

Detection method

Sample types

Reacts with

Results type

Sensitivity

Assay time

Assay Platform

Complementary Products

Sample Prep & Detection Kits

Secondary Antibodies

Primary Antibodies

Primary Antibodies

Product details

What's included?

Properties and storage information

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (9)

Product promise