AB207219

NFkB p52 Transcription Factor Assay Kit (Colorimetric)

Name: NFkB p52 Transcription Factor Assay Kit (Colorimetric) (ab207219) | Abcam
Brand: Abcam Limited
SKU: AB207219
Price: 5275 USD
Availability: InStock

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(1 Publication)

NFkB p52 Transcription Factor Assay Kit (Colorimetric) (ab207219) is a high throughput assay to quantify NFkB p52 activation in nuclear extracts.

View Alternative Names

LYT10, NFKB2, Nuclear factor NF-kappa-B p100 subunit, DNA-binding factor KBF2, H2TF1, Lymphocyte translocation chromosome 10 protein, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 2, Oncogene Lyt-10, Lyt10

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Functional Studies - NFkB p52 Transcription Factor Assay Kit (Colorimetric) (AB207219)

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Supplier Data

Functional Studies - NFkB p52 Transcription Factor Assay Kit (Colorimetric) (AB207219)

Nuclear extracts (10 μg/well) prepared from a variety of cell lines were tested for NFkB p52 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, Cell Lysate

Assay type

Semi-quantitative

Sensitivity

< 500 ng/well

Assay time

3h 30m

Assay Platform

Microplate reader

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Product details

NFkB p52 Transcription Factor Assay Kit (Colorimetric) (ab207219) is a high throughput assay to quantify NFkB p52 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 consensus binding site (5´ - GGGACTTTCC - 3´) has been immobilized onto a 96-well plate. Active NFkB present in nuclear or whole cell extracts specifically binds to the oligonucleotide. NFkB p52 is detected by a primary antibody that recognizes an epitope of NFkB p52 accessible only when NFkB 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 p52.

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?

{ "values": { "5x96Tests": { "sellingSize": "5 x 96 Tests", "publicAssetCode":"ab207219-5x96Tests", "assetComponentDetails": [ { "size":"5 x 1 Unit", "name":"96-well NFkB assay plate", "number":"AB207219-CMP03" }, { "size":"5 x 1 Unit", "name":"Plate sealer", "number":"AB207219-CMP12" }, { "size":"1 x 500 µL", "name":"Mutated oligonucleotide (10 pmol/µL)", "number":"AB207219-CMP06" }, { "size":"1 x 500 µL", "name":"Wild-type oligonucleotide (10 pmol/µL)", "number":"AB207219-CMP16" }, { "size":"1 x 11 mL", "name":"10X Antibody Binding Buffer", "number":"AB207219-CMP01" }, { "size":"1 x 55 mL", "name":"Developing Solution", "number":"AB207219-CMP09" }, { "size":"1 x 110 mL", "name":"10X Wash Buffer", "number":"AB207219-CMP02" }, { "size":"1 x 55 µL", "name":"NFkB p52 antibodies", "number":"AB207219-CMP11" }, { "size":"1 x 55 mL", "name":"Stop Solution", "number":"AB207219-CMP15" }, { "size":"1 x 500 µL", "name":"Herring sperm DNA", "number":"AB207219-CMP04" }, { "size":"1 x 55 µL", "name":"Anti-rabbit HRP-conjugated IgG", "number":"AB207219-CMP07" }, { "size":"1 x 50 mL", "name":"Binding Buffer", "number":"AB207219-CMP08" }, { "size":"1 x 200 µL", "name":"Raji nuclear extract (2.5 µg/µL)", "number":"AB207219-CMP14" }, { "size":"1 x 50 mL", "name":"Lysis Buffer", "number":"AB207219-CMP05" }, { "size":"1 x 500 µL", "name":"Dithiothreitol (DTT) (1 M)", "number":"AB207219-CMP10" }, { "size":"1 x 500 µL", "name":"Protease Inhibitor Cocktail", "number":"AB207219-CMP13" } ] }, "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab207219-1x96Tests", "assetComponentDetails": [ { "size":"1 x 100 µL", "name":"Wild-type oligonucleotide (10 pmol/µL)", "number":"AB207219-CMP16", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Plate sealer", "number":"AB207219-CMP12", "productcode":"" }, { "size":"1 x 1 Unit", "name":"96-well NFkB assay plate", "number":"AB207219-CMP03", "productcode":"" }, { "size":"1 x 100 µL", "name":"Mutated oligonucleotide (10 pmol/µL)", "number":"AB207219-CMP06", "productcode":"" }, { "size":"1 x 10 mL", "name":"Lysis Buffer", "number":"AB207219-CMP05", "productcode":"" }, { "size":"1 x 11 µL", "name":"NFkB p52 antibodies", "number":"AB207219-CMP11", "productcode":"" }, { "size":"1 x 2.2 mL", "name":"10X Antibody Binding Buffer", "number":"AB207219-CMP01", "productcode":"" }, { "size":"1 x 100 µL", "name":"Dithiothreitol (DTT) (1 M)", "number":"AB207219-CMP10", "productcode":"" }, { "size":"1 x 10 mL", "name":"Binding Buffer", "number":"AB207219-CMP08", "productcode":"" }, { "size":"1 x 100 µL", "name":"Herring sperm DNA", "number":"AB207219-CMP04", "productcode":"" }, { "size":"1 x 11 µL", "name":"Anti-rabbit HRP-conjugated IgG", "number":"AB207219-CMP07", "productcode":"" }, { "size":"1 x 100 µL", "name":"Protease Inhibitor Cocktail", "number":"AB207219-CMP13", "productcode":"" }, { "size":"1 x 11 mL", "name":"Developing Solution", "number":"AB207219-CMP09", "productcode":"" }, { "size":"1 x 40 µL", "name":"Raji nuclear extract (2.5 µg/µL)", "number":"AB207219-CMP14", "productcode":"" }, { "size":"1 x 11 mL", "name":"Stop Solution", "number":"AB207219-CMP15", "productcode":"" }, { "size":"1 x 22 mL", "name":"10X Wash Buffer", "number":"AB207219-CMP02", "productcode":"" } ] } } }

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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

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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 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. 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. In a non-canonical activation pathway, the MAP3K14-activated CHUK/IKKA homodimer phosphorylates NFKB2/p100 associated with RelB, inducing its proteolytic processing to NFKB2/p52 and the formation of NF-kappa-B RelB-p52 complexes. The NF-kappa-B heterodimeric RelB-p52 complex is a transcriptional activator. The NF-kappa-B p52-p52 homodimer is a transcriptional repressor. NFKB2 appears to have dual functions such as cytoplasmic retention of attached NF-kappa-B proteins by p100 and generation of p52 by a cotranslational processing. The proteasome-mediated process ensures the production of both p52 and p100 and preserves their independent function. p52 binds to the kappa-B consensus sequence 5'-GGRNNYYCC-3', located in the enhancer region of genes involved in immune response and acute phase reactions. p52 and p100 are respectively the minor and major form; the processing of p100 being relatively poor. Isoform p49 is a subunit of the NF-kappa-B protein complex, which stimulates the HIV enhancer in synergy with p65. In concert with RELB, regulates the circadian clock by repressing the transcriptional activator activity of the CLOCK-BMAL1 heterodimer.

See full target information NFKB2

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Publications (1)

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

eLife 10: PubMed34854378

2021

Single-cell-level protein analysis revealing the roles of autoantigen-reactive B lymphocytes in autoimmune disease and the murine model.

Applications

Unspecified application

Species

Unspecified reactive species

Takemichi Fukasawa,Ayumi Yoshizaki,Satoshi Ebata,Asako Yoshizaki-Ogawa,Yoshihide Asano,Atsushi Enomoto,Kiyoshi Miyagawa,Yutaka Kazoe,Kazuma Mawatari,Takehiko Kitamori,Shinichi Sato

View all publications

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Product promise

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Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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NFkB p52 Transcription Factor Assay Kit (Colorimetric)

Functional Studies - NFkB p52 Transcription Factor Assay Kit (Colorimetric) (AB207219)

Key facts

Detection method

Sample types

Assay type

Sensitivity

Assay time

Assay Platform

Product details

What's included?

Properties and storage information

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Product protocols

Target data

Publications (1)

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