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
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- Acta cirurgica brasileira 38:e3807232023Astragaloside IV alleviates stroke-triggered early brain injury by modulating neuroinflammation and ferroptosis via the Nrf2/HO-1 signaling pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesChunlei Zhang,Zhonghua Shi,Qinyi Xu,Jianqing He,Lei Chen,Zehua Lu,Qiaohua Huan,Yuhai Wang,Gang CuiPubMed 36995819
- Translational neuroscience 14:202202772023Omega-3 polyunsaturated fatty acids alleviate early brain injury after traumatic brain injury by inhibiting neuroinflammation and necroptosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYali Wu,Jing Zhang,Xiaoyan Feng,Wei JiaoPubMed 36895263
- International journal of molecular sciences 24:2023Activation of Endothelial Large Conductance Potassium Channels Protects against TNF-α-Induced Inflammation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTatiana Zyrianova,Kathlyn Zou,Benjamin Lopez,Andy Liao,Charles Gu,Riccardo Olcese,Andreas SchwingshacklPubMed 36835507
- Frontiers in nutrition 9:10602122023Surfeit folic acid, protein, and exercise modify oncogenic inflammatory biomarkers and fecal microbiota.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRebecca M Tuska,Sophia M Helm,C Foster Graf,Courtney James,Gabriel Kong,Leah T Stiemsma,David B Green,Susan Edgar HelmPubMed 36742002
- Mediators of inflammation 2022:11085692022Prevents Ulcerative Colitis by Promoting M2 Macrophage Polarization through Blocking the TLR4/MAPK/NF-B Pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJun Sun,Ping Zhao,Xufeng Ding,Fang Li,Jie Jiang,Hua Huang,Lijiang JiPubMed 36619207
- Acta cirurgica brasileira 37:e3706052022Cerebrolysin alleviates early brain injury after traumatic brain injury by inhibiting neuroinflammation and apoptosis via TLR signaling pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWeihong Lu,Zhonghua Zhu,Dongliang Shi,Xiaoyu Li,Jingzhi Luo,Xingzhi LiaoPubMed 36074398
- Acta cirurgica brasileira 37:e3706062022Ulinastatin alleviates early brain injury after intracerebral hemorrhage by inhibiting oxidative stress and neuroinflammation via ROS/MAPK/Nrf2 signaling pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesXi Wu,Wei Jiao,Junhui Chen,Yunna Tao,Jing Zhang,Yuhai WangPubMed 36074399
- Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 56:401-4172022Nootkatone Ameliorates Doxorubicin Induced Myocardial Injury through Modulation of NF-κB Signals and Oxidative Stress.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSuhail Al-Salam,Karthishwaran Kandhan,Manjusha Sudhadevi,Saeed TariqPubMed 36001774
- Frontiers in pharmacology 13:8872332022Carbocisteine as a Modulator of Nrf2/HO-1 and NFκB Interplay in Rats: New Inspiration for the Revival of an Old Drug for Treating Ulcerative Colitis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAmir Mohamed Abdelhamid,Mahmoud E Youssef,Simona Cavalu,Gomaa Mostafa-Hedeab,Amal Youssef,Sara T Elazab,Samar Ibrahim,Shady Allam,Rehab Mohamed Elgharabawy,Eman El-Ahwany,Noha A Amin,Ahmed Shata,Osama A Mohammed,Mahmoud Said Ibrahim Abdeldaiem,Ahmed Alhowail,Gaber El-Saber Batiha,Engy A El-Mahmoudy,Maram Attia,Alaa Allam,Mona Y Zaater,Mona M Osman,Manar Nader,Aya Taha,Nada Abul Makarem,Sameh SaberPubMed 35754464
- Acta cirurgica brasileira 37:e3703012022Ulinastatin alleviates early brain injury after intracerebral hemorrhage by inhibiting necroptosis and neuroinflammation via MAPK/NF-κB signaling pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLi Wang,Wei Jiao,Jiayu Wu,Jing Zhang,Min Tang,Yang ChenPubMed 35584533
Key facts
- Detection method
- Colorimetric
- Sample types
- Tissue Homogenate, Cell Lysate
- Assay type
- Semi-quantitative
- Reactive species
- Mouse, Human
- Assay time
- 1h 30m
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
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Target data
Function
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).
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
What's included?
Recommended products
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
Key facts
- Detection method
- Colorimetric
- Sample types
- Tissue Homogenate, Cell Lysate
- Assay type
- Semi-quantitative
- Reactive species
- Mouse, Human
- Assay time
- 1h 30m
- Assay Platform
- Microplate
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample (pS536) | n 6 | C.V. 3.4 |
Sample Overall | n 6 | C.V. 2.8 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample (pS536) | n 3 | C.V. 10.3 |
Sample Overall | n 3 | C.V. 8.7 |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
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
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
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.
- Associated diseases and disorders
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.
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4 product images
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)
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)
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)
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).
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