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AB176647

NF kappaB p65 (pS536) ELISA Kit

5

(2 Reviews)

|

(22 Publications)

NF kappaB p65 (pS536) ELISA Kit is a single-wash 90-min Simplestep used to quantify NF kappaB p65 (pS536) with a sensitivity of 5 µg/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Cited in over 15 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

4 Images
Sandwich ELISA - NF kappaB p65 (pS536) ELISA Kit (AB176647)
  • sELISA

Supplier Data

Sandwich ELISA - NF kappaB p65 (pS536) ELISA Kit (AB176647)

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 NFκB p65 (pS536) are normalized and plotted.

Sandwich ELISA - NF kappaB p65 (pS536) ELISA Kit (AB176647)
  • sELISA

Supplier Data

Sandwich ELISA - NF kappaB p65 (pS536) ELISA Kit (AB176647)

Typical cell lysate dilution series.

Example of a typical NFκB p65 (pS536) cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - NF kappaB p65 (pS536) ELISA Kit (AB176647)
  • sELISA

Supplier Data

Sandwich ELISA - NF kappaB p65 (pS536) ELISA Kit (AB176647)

Comparison of NF kappaB p65 (pS536) expression in different cell lines.

Cell line analysis for Total NFκB 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).

Sandwich ELISA - NF kappaB p65 (pS536) ELISA Kit (AB176647)
  • sELISA

Supplier Data

Sandwich ELISA - NF kappaB p65 (pS536) ELISA Kit (AB176647)

NF kappaB p65 (pS536) phosphorylation in response to TNFα treatment.

Induction of NFκB 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 NFκB p65 (pS536) are plotted and compared against total NFκB p65 protein levels. Comparative NFκB p65 (pS536) and NFκB p65 (Total) data also shown by Western Blot.

Key facts

Detection method

Colorimetric

Sample types

Tissue Homogenate, Cell Lysate

Reacts with

Mouse, Human

Assay type

Semi-quantitative

Sensitivity

= 5 µg/mL

Range

6 - 600 µg/mL

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 NFκB p65 (pS536) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of NFκB p65 (pS536) 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.

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": "HeLa Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "10.3" }, { "reproducibilityType": "Intra", "sample": "HeLa Extract", "replicates": 6, "mean": null, "standardDeviation": null, "coefficientOfVariability": "3.4" } ]

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 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.

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. Beside 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 (22)

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

Frontiers in pharmacology 16:1555079 PubMed40144659

2025

Vortioxetine alleviates motor, cognitive and emotional disorders in post-stroke rats by regulating the TLR-2/NF-B pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Ziqiang Dong,Zhihui Dong,Lili Xu,Jinfeng Zhang,Lin Li,Rongjuan Wang,Xiaoyan Huang,Zhengqiang Zou

Inflammopharmacology 32:1839-1853 PubMed38581641

2024

Terminalia ferdinandiana Exell. extracts reduce pro-inflammatory cytokine and PGE secretion, decrease COX-2 expression and down-regulate cytosolic NF-κB levels.

Applications

Unspecified application

Species

Unspecified reactive species

Ian E Cock

Scientific reports 13:11899 PubMed37488162

2023

Anandamide modulates WNT-5A/BCL-2, IP3/NFATc1, and HMGB1/NF-κB trajectories to protect against mercuric chloride-induced acute kidney injury.

Applications

Unspecified application

Species

Unspecified reactive species

Dalaal M Abdallah,Mahmoud M Kamal,Nour Eldin S Aly,Hanan S El-Abhar

Disease models & mechanisms 16: PubMed37021539

2023

Vemurafenib improves muscle histopathology in a mouse model of LAMA2-related congenital muscular dystrophy.

Applications

Unspecified application

Species

Unspecified reactive species

Ariany Oliveira-Santos,Marisela Dagda,Jennifer Wittmann,Robert Smalley,Dean J Burkin

Cell journal 25:17-24 PubMed36680480

2023

Nuclear Factor Kappa-B Protein Levels in Sperm of Obese Men with and without Diabetes; Cellular Approach in Male Infertility.

Applications

Unspecified application

Species

Unspecified reactive species

Shima Abbasihormozi,Azam Kouhkan,Abdolhossein Shaverdi,Mohammad Ali Sadighi Gilani,Vahab Babapour,Amir Niasari Naslji,Vahid Akbarinehad,AliReza Alizadeh

Nutrients 14: PubMed35956298

2022

A Comparison of the Impact of Restrictive Diets on the Gastrointestinal Tract of Mice.

Applications

Unspecified application

Species

Unspecified reactive species

András Gregor,Laura Huber,Sandra Auernigg-Haselmaier,Felix Sternberg,Magdalena Billerhart,Andreas Dunkel,Veronika Somoza,Manfred Ogris,Barbara Kofler,Valter D Longo,Jürgen König,Kalina Duszka

Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics 18:2664-2681 PubMed34664178

2021

Crosstalk Among NLRP3 Inflammasome, ETR Signaling, and miRNAs in Stress-Induced Depression-Like Behavior: a Modulatory Role for SGLT2 Inhibitors.

Applications

Unspecified application

Species

Unspecified reactive species

Radwa N Muhammad,Lamiaa A Ahmed,Rania M Abdul Salam,Kawkab A Ahmed,Amina S Attia

Molecular medicine reports 23: PubMed33537800

2021

Diagnostic value of microRNA‑125b in peripheral blood mononuclear cells for pulmonary tuberculosis.

Applications

Unspecified application

Species

Unspecified reactive species

Xiaolu Sun,Kai Liu,Xinhong Wang,Tianhua Zhang,Xiaomou Li,Yan Zhao

Molecules (Basel, Switzerland) 26: PubMed33440679

2021

Zeaxanthin-Rich Extract from Superfood Selectively Modulates the Cellular Adhesion and MAPK Signaling in Melanoma versus Normal Skin Cells In Vitro.

Applications

Unspecified application

Species

Unspecified reactive species

Diana Cenariu,Eva Fischer-Fodor,Adrian Bogdan Țigu,Andrea Bunea,Piroska Virág,Maria Perde-Schrepler,Vlad-Alexandru Toma,Andrei Mocan,Ioana Berindan-Neagoe,Adela Pintea,Gianina Crișan,Mihai Cenariu,Alma Maniu

Scientific reports 10:14693 PubMed32895407

2020

Activated ROCK/Akt/eNOS and ET-1/ERK pathways in 5-fluorouracil-induced cardiotoxicity: modulation by simvastatin.

Applications

Unspecified application

Species

Unspecified reactive species

Radwa Nasser Muhammad,Nada Sallam,Hanan Salah El-Abhar
View all publications
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