Recombinant Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)
![Western blot - Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)](https://www.abcam.com/ps/products/183/ab183559/Images/ab183559-456775-anti-nf-kb-p65-phospho-s276-antibody-epr17622-western-blot-hela-human.jpg "Western blot - Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17622] to NF-kB p65 (phospho S276)
- Suitable for: ICC/IF, IP, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-NF-kB p65 (phospho S276) antibody [EPR17622]
See all NF-kB p65 primary antibodies -
Description
Rabbit monoclonal [EPR17622] to NF-kB p65 (phospho S276) -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IP, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Lysates of HeLa cells treated with Calyculin A (100ng/ml, 30min), then TNF-a (20ng/ml, 5min). ICC/IF: HeLa cells treated with Calyculin A (100ng/ml, 10min), then TNF-a (20ng/ml, 5min). IP: HeLa whole cell lysate treated with Calyculin A (100ng/ml, 10min), then TNF-a (20ng/ml, 5min).
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17622 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab183559 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF |
1/100.
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| IP |
1/40.
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| WB |
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 60 kDa).
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| Notes |
|---|
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ICC/IF
1/100. |
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IP
1/40. |
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WB
1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 60 kDa). |
Target
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Function
NF-kappa-B is a pleiotropic transcription factor which is present in almost all cell types and is involved in many biological processed 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 and the heterodimeric p65-p50 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. 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. NF-kappa-B heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kappa-B p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. The inhibitory effect of I-kappa-B upon NF-kappa-B the cytoplasm is exerted primarily through the interaction with p65. p65 shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. -
Sequence similarities
Contains 1 RHD (Rel-like) domain. -
Domain
the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors. -
Post-translational
modificationsUbiquitinated, leading to its proteasomal degradation. Degradation is required for termination of NF-kappa-B response.
Monomethylated at Lys-310 by SETD6. Monomethylation at Lys-310 is recognized by the ANK repeats of EHMT1 and promotes the formation of repressed chromatin at target genes, leading to down-regulation of NF-kappa-B transcription factor activity. Phosphorylation at Ser-311 disrupts the interaction with EHMT1 without preventing monomethylation at Lys-310 and relieves the repression of target genes.
Phosphorylation at Ser-311 disrupts the interaction with EHMT1 and promotes transcription factor activity (By similarity). Phosphorylation on Ser-536 stimulates acetylation on Lys-310 and interaction with CBP; the phosphorylated and acetylated forms show enhanced transcriptional activity.
Reversibly acetylated; the acetylation seems to be mediated by CBP, the deacetylation by HDAC3. Acetylation at Lys-122 enhances DNA binding and impairs association with NFKBIA. Acetylation at Lys-310 is required for full transcriptional activity in the absence of effects on DNA binding and NFKBIA association. Acetylation can also lower DNA-binding and results in nuclear export. Interaction with BRMS1 promotes deacetylation of 'Lys-310'. -
Cellular localization
Nucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B). Colocalized with RELA in the nucleus upon TNF-alpha induction. - Information by UniProt
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Database links
- Entrez Gene: 5970 Human
- Omim: 164014 Human
- SwissProt: Q04206 Human
- Unigene: 502875 Human
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Alternative names
- Avian reticuloendotheliosis viral (v rel) oncogene homolog A antibody
- MGC131774 antibody
- NF kappa B p65delta3 antibody
see all
Images
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Western blot - Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)All lanes : Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559) at 1/1000 dilution
Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa whole cell lysate treated with 100ng/ml Calyculin A ab141784 for 30 minutes, then treated with 20ng/ml TNF-a for 5 minutes
Lane 3 : HeLa whole cell lysate treated with 100ng/ml Calyculin A ab141784 for 30 minutes, then treated with 20ng/ml TNF-a for 5 minutes, then treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 60 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesNegative control: Anti-NF-kB p65 antibody, ab32536
Blocking/Dilution buffer: 5% NFDM/TBST.
Phosphorylation of NF-kB p65 at S276 can be induced by TNF-a treatment according to the literature (PMID: 21795584).
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![Immunocytochemistry/ Immunofluorescence - Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)](https://www.abcam.com/ps/products/183/ab183559/Images/ab183559-269448-anti-nf-kb-p65-phospho-s276-antibody-epr17622-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling NF-kB p65 (phospho S276) with ab183559 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
The expression increased on HeLa cells after treatment with Calyculin A (ab141784 100ng/ml, 10min) then TNF-a (20ng/ml, 5min).
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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![Immunoprecipitation - Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)](https://www.abcam.com/ps/products/183/ab183559/Images/ab183559-269445-anti-nf-kb-p65-phospho-s276-antibody-epr17622-immunoprecipitation.jpg)
Immunoprecipitation - Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)NF-kB p65 (phospho S276) was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates, treated with 100ng/ml Calyculin A (ab141784) for 10min, then 20ng/ml TNA-a for 5min, with ab183559 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab183559 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa whole cell lysate treated with 100ng/ml CA for 10min, then 20ng/ml TNA-a for 5min, 10 μg (Input).
Lane 2: ab183559 IP in HeLa whole cell lysate treated with 100ng/ml CA for 10min, then 20ng/ml TNA-a for 5min.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab183559 in HeLa whole cell lysate treated with 100ng/ml CA for 10min, then 20ng/ml TNA-a for 5min.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
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![Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)](https://www.abcam.com/ps/products/183/ab183559/Images/ab183559-6-benefits-of-recombinant-antibodies.png)
Anti-NF-kB p65 (phospho S276) antibody [EPR17622] (ab183559)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (12)
ab183559 has been referenced in 12 publications.
- Xiao X et al. DUOX2 participates in skin aging induced by UVB in HSF2 cells by activating NF-?B signaling. Exp Ther Med 21:157 (2021). PubMed: 33456524
- Zhao J et al. Lipoxin A4 promotes autophagy and inhibits overactivation of macrophage inflammasome activity induced by Pg LPS. J Int Med Res 49:300060520981259 (2021). PubMed: 33528285
- Xu J et al. The RNA helicase DDX5 promotes viral infection via regulating N6-methyladenosine levels on the DHX58 and NF?B transcripts to dampen antiviral innate immunity. PLoS Pathog 17:e1009530 (2021). PubMed: 33909701
- Ling P et al. miR-1184 regulates inflammatory responses and cell apoptosis by targeting TRADD in an LPS-induced cell model of sepsis. Exp Ther Med 21:630 (2021). PubMed: 33936286
- Yang Z et al. A Novel Mechanical-Based Injective Hydrogel for Treatment with Aromatase Inhibitors Caused Joint Inflammation via the NF-?B Pathway. ACS Omega 6:10242-10249 (2021). PubMed: 34056178