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AB176663

NF kappaB p65 (pS536 + Total) ELISA Kit

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(31 Publications)

NF kappaB p65 (pS536 + Total) ELISA Kit is a single-wash 90-min Simplestep used to detect NF kappaB p65 (pS536 + Total). The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Semi-quantitative Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Cited in over 25 citations

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

5 Images
ELISA - NF kappaB p65 (pS536 + Total) ELISA Kit (AB176663)
  • ELISA

Supplier Data

ELISA - NF kappaB p65 (pS536 + Total) ELISA Kit (AB176663)
Sandwich ELISA - NF kappaB p65 (pS536 + Total) ELISA Kit (AB176663)
  • sELISA

Supplier Data

Sandwich ELISA - NF kappaB p65 (pS536 + Total) ELISA Kit (AB176663)

Typical cell lysate dilution series.

Example of a typical NFĸB p65 (pS536) and NFĸB p65 (Total) cell lysate dilution series. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - NF kappaB p65 (pS536 + Total) ELISA Kit (AB176663)
  • sELISA

Supplier Data

Sandwich ELISA - NF kappaB p65 (pS536 + Total) ELISA Kit (AB176663)

Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1X Cell Extraction Buffer PTR. Data from duplicate measurements of NFĸB p65 (pS536) are normalized and plotted.

Sandwich ELISA - NF kappaB p65 (pS536 + Total) ELISA Kit (AB176663)
  • sELISA

Supplier Data

Sandwich ELISA - NF kappaB p65 (pS536 + Total) ELISA Kit (AB176663)

NFkB p65 (pS536) phosphorylation in response to TNFα treatment.

Induction of NFĸB p65 (pS536) phosphorylation in MCF-7 cells in response to TNFα treatment. HeLa cells were cultured in 96-well tissue culture plates, and treated (10 min) with a dose-range of TNFα before cell lysis. Data from quadruplicate measurements of NFĸB p65 (pS536) are plotted and compared against total NFĸB p65 protein levels. Comparative NFĸB p65 (pS536) and NFĸB p65 (Total) data also shown by Western Blot.

Sandwich ELISA - NF kappaB p65 (pS536 + Total) ELISA Kit (AB176663)
  • sELISA

Supplier Data

Sandwich ELISA - NF kappaB p65 (pS536 + Total) ELISA Kit (AB176663)

Comparison of NFkB p65 expression in different cell lines.

Cell line analysis for Total NFĸB p65 from 200 μg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).

Key facts

Detection method

Colorimetric

Sample types

Tissue Homogenate, Cell Lysate

Reacts with

Mouse, Human

Results type

Semi-Quantitative

Assay time

1h 30m

Assay Platform

Microplate

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "ELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

Abcam's NFkB p65 (pS536) and NFkB p65 (Total) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of NFkB p65 (pS536) and Total NFkB p65 protein in Human and mouse cells.

The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

As of October 2019, this kit was reformulated with new antibodies to maintain continued long term supply.

Estimated sensitivity: Phospho-NF-κB p65 (Ser536): 5 μg/mL (tested with HeLa), Total NF-κB p65: 5 μg/mL (tested with HeLa)
Range: Phospho-NF-κB p65 (Ser536): 6-600 μg/mL, Total NF-κB p65: 6-600 μg/mL

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Precision

[ { "reproducibilityType": "Inter", "sample": "(pS536)", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "10.3" }, { "reproducibilityType": "Inter", "sample": "Overall", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "8.7" }, { "reproducibilityType": "Intra", "sample": "(pS536)", "replicates": 6, "mean": null, "standardDeviation": null, "coefficientOfVariability": "3.4" }, { "reproducibilityType": "Intra", "sample": "Overall", "replicates": 6, "mean": null, "standardDeviation": null, "coefficientOfVariability": "2.8" } ]

What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Supplementary information

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

NF-kB p65 also known as RelA is a transcription factor with a molecular mass of around 65 kDa. It is a vital component of the NF-kB family typically found in the cytoplasm in its inactive form. Upon activation it translocates to the nucleus where it regulates the expression of target genes. NF-kB p65 is ubiquitously expressed in various cell types including immune cells and epithelial cells.
Biological function summary

NF-kB p65 plays a central role in regulating immune response and inflammation. It forms a complex with proteins such as p50 to bind DNA and modulate gene transcription. This complex is essential for cellular responses to stress and infection influencing processes such as cell proliferation and survival. It also interacts with other transcription factors and cofactors to fine-tune gene regulation.

Pathways

NF-kB p65 is integral to the canonical NF-kB signaling pathway which gets activated by signals like cytokines and bacterial products. The protein family includes several related proteins such as IκB that inhibit NF-kB activity until signal-induced degradation allows NF-kB to enter the nucleus. Another related pathway is the MAPK pathway which can interact with NF-kB signaling to coordinate cellular stress responses.

NF-kB p65 is implicated in conditions such as cancer and inflammatory diseases. Its dysregulation can lead to chronic inflammation contributing to tumor progression. Inflammatory disorders like rheumatoid arthritis also show abnormal NF-kB activity contributing to disease pathology. Additionally interactions with proteins like IKK contribute significantly to these diseases altering NF-kB signaling and gene expression patterns.

Product protocols

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 phospho S536

Publications (31)

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Food science & nutrition 12:9566-9576 PubMed39619973

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Anti-Inflammatory Effects of Fermented and Aged Mountain-Cultivated Ginseng Sprouts via Suppression of MAPK-NF-κB Pathway in Lipopolysaccharide-Stimulated RAW264.7 Macrophages.

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Dang Long Cao,Min-Seok Woo,Eun-Jin Kim,Byeonggyu Ahn,Anjas Happy Prayoga,Sang Soo Kang,Kye Man Cho,Dawon Kang

Journal of diabetes investigation 15:1365-1376 PubMed39171660

2024

Calpeptin improves the cognitive function in Alzheimer's disease-like complications of diabetes mellitus rats by regulating TXNIP/NLRP3 inflammasome.

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Luyao Qiao,Shouqin Yi,Tianpei Li,Xin Pan,Gege Wang,Xu Liu,Min Li,Jun Min,Huahui Le,Zhenyu Tang

American journal of translational research 16:1102-1117 PubMed38715815

2024

Semaglutide alleviates early brain injury following subarachnoid hemorrhage by suppressing ferroptosis and neuroinflammation via SIRT1 pathway.

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Lei Chen,Haiting Xu,Chunlei Zhang,Jianqing He,Yuhai Wang

Acta cirurgica brasileira 38:e380723 PubMed36995819

2023

Astragaloside IV alleviates stroke-triggered early brain injury by modulating neuroinflammation and ferroptosis via the Nrf2/HO-1 signaling pathway.

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Chunlei Zhang,Zhonghua Shi,Qinyi Xu,Jianqing He,Lei Chen,Zehua Lu,Qiaohua Huan,Yuhai Wang,Gang Cui

Translational neuroscience 14:20220277 PubMed36895263

2023

Omega-3 polyunsaturated fatty acids alleviate early brain injury after traumatic brain injury by inhibiting neuroinflammation and necroptosis.

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Yali Wu,Jing Zhang,Xiaoyan Feng,Wei Jiao

International journal of molecular sciences 24: PubMed36835507

2023

Activation of Endothelial Large Conductance Potassium Channels Protects against TNF-α-Induced Inflammation.

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Tatiana Zyrianova,Kathlyn Zou,Benjamin Lopez,Andy Liao,Charles Gu,Riccardo Olcese,Andreas Schwingshackl

Frontiers in nutrition 9:1060212 PubMed36742002

2023

Surfeit folic acid, protein, and exercise modify oncogenic inflammatory biomarkers and fecal microbiota.

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Rebecca M Tuska,Sophia M Helm,C Foster Graf,Courtney James,Gabriel Kong,Leah T Stiemsma,David B Green,Susan Edgar Helm

Mediators of inflammation 2022:1108569 PubMed36619207

2022

Prevents Ulcerative Colitis by Promoting M2 Macrophage Polarization through Blocking the TLR4/MAPK/NF-B Pathway.

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Jun Sun,Ping Zhao,Xufeng Ding,Fang Li,Jie Jiang,Hua Huang,Lijiang Ji

Acta cirurgica brasileira 37:e370606 PubMed36074399

2022

Ulinastatin alleviates early brain injury after intracerebral hemorrhage by inhibiting oxidative stress and neuroinflammation via ROS/MAPK/Nrf2 signaling pathway.

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Species

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Xi Wu,Wei Jiao,Junhui Chen,Yunna Tao,Jing Zhang,Yuhai Wang

Acta cirurgica brasileira 37:e370605 PubMed36074398

2022

Cerebrolysin alleviates early brain injury after traumatic brain injury by inhibiting neuroinflammation and apoptosis via TLR signaling pathway.

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Species

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Weihong Lu,Zhonghua Zhu,Dongliang Shi,Xiaoyu Li,Jingzhi Luo,Xingzhi Liao
View all publications
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