AB239882

Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free

RabMAb
Recombinant
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(56 Publications)

Rabbit Recombinant Monoclonal NF-kB p65 phospho S536 antibody. Carrier free. Suitable for IP, Dot, WB and reacts with Human, Mouse, Rat, Synthetic peptide samples. Cited in 56 publications.

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

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Immunoprecipitation - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

ab76302 at 1/30 immunoprecipitating NF-kB p65 (phospho S536) in Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha.

Lane 1 (input) : Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha (10μg)

Lane 2 (+) : ab76302 + Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha.

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab76302 in Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha.

For western blotting, ab76302 at 1/500 dilution (0.95 μg/ml) and VeriBlot for IP Detection Reagent (HRP)(ab131366) at 1/1000 dilution were used.

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76302).

All lanes:

Immunoprecipitation - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] (<a href='/en-us/products/primary-antibodies/nf-kb-p65-phospho-s536-antibody-ep2294y-ab76302'>ab76302</a>)

Predicted band size: 60 kDa

Observed band size: 65 kDa

false

Unknown

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

All lanes:

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] (<a href='/en-us/products/primary-antibodies/nf-kb-p65-phospho-s536-antibody-ep2294y-ab76302'>ab76302</a>) at 1/1000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) treated with Calyculin A and TNF-alpha whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 60 kDa

Observed band size: 65 kDa

false

Unknown

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

All lanes:

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (ab239882) at 1/1000 dilution

Lane 1:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 15 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) treated with 100nM Calyculin A for 30 minutes whole cell lysate at 15 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast) treated with 100nM Calyculin A for 30 minutes, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 60 kDa

Observed band size: 65 kDa

false

Unknown

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

All lanes:

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] (<a href='/en-us/products/primary-antibodies/nf-kb-p65-phospho-s536-antibody-ep2294y-ab76302'>ab76302</a>) at 1/1000 dilution

Lane 1:

C6 (Rat glial tumor glial cell) whole cell lysate at 15 µg

Lane 2:

C6 (Rat glial tumor glial cell) treated with 100nM Calyculin A for 30 minutes whole cell lysate at 15 µg

Lane 3:

C6 (Rat glial tumor glial cell) treated with 100nM Calyculin A for 30 minutes, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 60 kDa

Observed band size: 65 kDa

false

Unknown

Dot Blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

Dot blot analysis of INF- kB p65 (phospho S536) phospho peptide (Lane 1) and NF- kB p65 non-phospho peptide (Lane 2) labeling NF-kB p65 (phospho S536) with ab76302 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG) (H+L) at 1/100 000 was used as the secondary antibody.

Blocking and diluting buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76302).

Unconjugated

Anti-NF-kB p65 (phospho S536) antibody [EP2294Y]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EP2294Y

Isotype

IgG

Carrier free

Yes

Reacts with

Mouse, Rat, Human

Applications

Dot, IP, WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

Stimulation may be required to allow detection of the phosphorylated protein.
We recommend using NIH/3T3 (Mouse embryonic fibroblast) treated with 100nM Calyculin A for 30 minutes as a positive control.

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Reactivity data

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Product details

ab239882 is the carrier-free version of ab76302.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

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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

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

Visit the General protocols
Visit the Troubleshooting

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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 pS536

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Publications (56)

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

Journal of gastrointestinal oncology 16:1443-1460 PubMed40950333

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NKAP overexpression promotes gastric cancer immune escape by inducing IL-10 secretion from mature dendritic cells during anti-PD-L1 therapy.

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Species

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Bingna Yan,Pei Zhang,Qianhe Zu,Shan Liu,Pan Wang,Yaning Wei,Jialin Liu,Shun Zhang,Yanan Wang,Zhe Han

Foods (Basel, Switzerland) 14: PubMed40870671

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Sprouted Black Quinoa Extract Alleviates Heat Stress-Induced Liver Injury in Rats by Activating Nrf2 Signaling and Suppressing the NF-κB/NLRP3 Inflammasome Pathway.

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Species

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Theranostics 15:5719-5737 PubMed40365290

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A bifunctional fusion membrane-based biocompatible nanovaccine to potentiate cancer immunotherapy.

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Wei Fu,Xing Cai,Jinru Yang,Lian Yang,Yaoyu Pan,Zhan Tuo

Molecular medicine reports 31: PubMed40084693

2025

Exosomes derived from baicalin‑pretreated mesenchymal stem cells mitigate atherosclerosis by regulating the SIRT1/NF‑κB signaling pathway.

Applications

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Species

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Xiaochun Yang,Wei Wu,Weitian Huang,Junfeng Fang,Yunli Chen,Xiaoyan Chen,Xiaolan Lin,Yanbin He

PeerJ 13:e18876 PubMed39897488

2025

Dihydromyricetin inhibits injury caused by ischemic stroke through the lncRNA SNHG17/miR-452-3p/CXCR4 axis.

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Species

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Jiacheng Xie,Qiuyue Yang,Xueliang Zeng,Qi Zeng,Hai Xiao

Microbial biotechnology 18:e70083 PubMed39825784

2025

Akkermansia muciniphila Mediated the Preventive Effect of Disulfiram on Acute Liver Injury via PI3K/Akt Pathway.

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Ruonan Zhang,Xuewei Sun,Han Lu,Xinrui Zhang,Mingyan Zhang,Xuewen Ji,Xinyi Yu,Chengliang Tang,Zihan Wu,Yinghua Mao,Jin Zhu,Minjun Ji,Zhan Yang

Drug development research 85:e70033 PubMed39660541

2024

Heme Oxygenase-1 Overexpression Activates the IRF1/DRP1 Signaling Pathway to Promote M2-Type Polarization of Spinal Cord Microglia.

Applications

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Species

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Wenping Lin,Ziming Cai,Jinzhu Liang,Ping Miao,Ye Ruan,Pian Li,Shuhui Lin,He Tian,Qinghe Yu,Xu He

Scientific reports 14:23074 PubMed39367100

2024

Quercetin as a therapeutic agent activate the Nrf2/Keap1 pathway to alleviate lung ischemia-reperfusion injury.

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Species

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Mohammad Yousefi Zardak,Fatemeh Keshavarz,Ali Mahyaei,Morteza Gholami,Fatemeh Sadat Moosavi,Elham Abbasloo,Farzaneh Abdollahi,Maryam Hossein Rezaei,Elham Madadizadeh,Nasrin Soltani,Fatemeh Bejeshk,Niyan Salehi,Fahimeh Rostamabadi,Fatemeh Bagheri,Mahla Jafaraghae,Mahdiyeh Ranjbar Zeydabadi,Meraj Baghgoli,Gholamreza Sepehri,Mohammad Abbas Bejeshk

Frontiers in pharmacology 15:1449985 PubMed39263577

2024

Protective effects of insulin on dry eye syndrome via TLR4/NF-κB pathway: based on network pharmacology and experiments validation.

Applications

Unspecified application

Species

Unspecified reactive species

Xiuxiu Yuan,Yu Zhang,Siyi Wang,Zhiyu Du

Heliyon 10:e34596 PubMed39114061

2024

A classic prescription alleviates inflammation in CUMS model mice via modulating MYDGF/MAP4K4/NF-κB signaling pathway, verified through UPLC-HRMS and proteomics analysis.

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Species

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Ruolan Huang,Shenglan Gong,Bocheng Xiong,Xifei Yang,Chongyang Chen,Wei Song,Ruodai Wu,Li Yang,Jia Yin,Mingtai Chen

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

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

For licensing inquiries, please contact partnerships@abcam.com

Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free

Immunoprecipitation - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

Western blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

Dot Blot - Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] - BSA and Azide free (AB239882)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Specificity

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Secondary Antibodies

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Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

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 (56)

Product promise