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AB207217

NFkB p50 Transcription Factor Assay Kit (Colorimetric)

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(3 Publications)

NFkB p50 Transcription Factor Assay Kit (Colorimetric) (ab207217) is a high throughput assay to quantify NFkB p50 activation in nuclear extracts.
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Functional Studies - NFkB p50 Transcription Factor Assay Kit (Colorimetric) (AB207217)
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Supplier Data

Functional Studies - NFkB p50 Transcription Factor Assay Kit (Colorimetric) (AB207217)

Nuclear extracts were assayed for NFkB p50 activity using ab207217.

Nuclear extracts prepared from untreated HeLa cells (Light gray), HeLa cells treated with TNF-α (Dark gray), untreated Jurkat cells (White), Jurkat cells treated with PMA and calcium ionophore (CI) (Black) and Raji cells (Black dots on white) were assayed at 10 μg/well for NFkB p50 activity using ab207217. Data shown are the results from wells assayed in duplicate. These results are provided for demonstration only.

Key facts

Detection method

Colorimetric

Sample types

Nuclear Extracts, Cell Lysate

Reacts with

Mouse, Human

Assay type

Semi-quantitative

Sensitivity

< 500 ng/well

Assay time

3h 30m

Assay Platform

Microplate reader

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Product details

NFkB p50 Transcription Factor Assay Kit (Colorimetric) (ab207217) is a high throughput assay to quantify NFkB p50 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 p50 consensus binding site (5´ - GGGACTTTCC - 3´) has been immobilized onto a 96-well plate. Active NFkB p50 present in nuclear or whole cell extracts specifically binds to the oligonucleotide. NFkB p50 is detected by a primary antibody that recognizes an epitope of NFkB p50 accessible only when the protein is activated and bound to its target DNA. An HRP-conjugated secondary antibody provides sensitive colorimetric readout at OD 450 nm. This product detects human and mouse NFkB p50.

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 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 IkBγ 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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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

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Publications (3)

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

Journal of biochemical and molecular toxicology 36:e23119 PubMed35678308

2022

The lncARSR/PTEN/Akt/nuclear factor-kappa B feedback regulatory loop contributes to doxorubicin resistance in hepatocellular carcinoma.

Applications

Unspecified application

Species

Unspecified reactive species

Yaling Li,Jun Li,Hongyan Chen,Jiting Wang,Linhan Jiang,Xiaoxia Tan

Molecular oncology 14:3211-3233 PubMed33040438

2020

Long noncoding RNA LINC01578 drives colon cancer metastasis through a positive feedback loop with the NF-κB/YY1 axis.

Applications

Unspecified application

Species

Unspecified reactive species

Jia Liu,Yang Zhan,Jiefu Wang,Junfeng Wang,Jiansheng Guo,Dalu Kong

Frontiers in microbiology 9:385 PubMed29568285

2018

Activates Hypoxia Inducible Factor-1α and miR-210 for Survival in Macrophages by Downregulation of NF-κB Mediated Pro-inflammatory Immune Response.

Applications

Unspecified application

Species

Unspecified reactive species

Vinod Kumar,Ajay Kumar,Sushmita Das,Ashish Kumar,Kumar Abhishek,Sudha Verma,Abhishek Mandal,Rakesh K Singh,Pradeep Das
View all publications
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions
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