Anti-NF-kB p65 (acetyl K310) antibody (ab19870)
Key features and details
- Rabbit polyclonal to NF-kB p65 (acetyl K310)
- Suitable for: WB, IP, Dot blot, ICC
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-NF-kB p65 (acetyl K310) antibody
See all NF-kB p65 primary antibodies -
Description
Rabbit polyclonal to NF-kB p65 (acetyl K310) -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, Dot blot, ICCmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide corresponding to Human NF-kB p65 aa 300-400 (internal sequence) conjugated to keyhole limpet haemocyanin.
(Peptide available asab20612) -
Positive control
- This antibody gave a positive signal with acetylated p65 protein and Rat lung tissue lysate.
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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Assay kits
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ChIP Related Products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
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Related Products
Applications
Our Abpromise guarantee covers the use of ab19870 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 |
|---|---|---|
| WB | Use a concentration of 2.5 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa). Collaborator data suggests that immunoprecipitation of this antibody prior to Western blotting is required to obtain the best results (see images) | |
| IP | Use a concentration of 2.5 µg/ml. | |
| Dot blot | Use at an assay dependent concentration. | |
| ICC | Use at an assay dependent concentration. In ICC/IF ab19870 recognizes various acetylated nuclear protein(s), as the signal is also observed in control cells; the signal in ICC is HDACi-dependent. |
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
- Entrez Gene: 19697 Mouse
- Entrez Gene: 309165 Rat
- Omim: 164014 Human
- SwissProt: Q04206 Human
- SwissProt: Q04207 Mouse
- Unigene: 502875 Human
- Unigene: 249966 Mouse
see all -
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 (acetyl K310) antibody (ab19870)This image is courtesy of Prof. Michael Hottiger's Lab, University of ZurichLanes 1-2 : Coomassie stain
Lanes 3-4 : Anti-NF-kB p65 (acetyl K310) antibody (ab19870) at 1/500 dilution
Lanes 1 & 3 : Acetylated p65 protein
Lanes 2 & 4 : K310R mutant protein
Lysates/proteins at 2 µg per lane.
Secondary
Lanes 3-4 : IRDye® 800CW Goat anti-Rabbit IgG (H + L) at 1/15000 dilution
Predicted band size: 65 kDaObserved band size: 75kDa
The p65 band runs higher in this blot because the protein contains a myc-tag.
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Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)This image is courtesy of an Abreview submitted by Christian MarxAll lanes : Anti-NF-kB p65 (acetyl K310) antibody (ab19870) at 1/1000 dilution
Lane 1 : HCT116 whole cell lysate treated with DMSO for 24 hrs (control)
Lane 2 : HCT116 whole cell lysate treated with 2 µM SAHA for 24 hrs
Lane 3 : MEF whole cell lysate treated with DMSO for 24 hrs (control)
Lane 4 : MEF whole cell lysate treated with 2 µM SAHA for 24 hrs
Lysates/proteins at 60 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 65 kDa
Observed band size: 65 kDa
Additional bands at: 140 kDa (possible non-specific binding), 15 kDa (possible non-specific binding), 40 kDa (possible non-specific binding), 45 kDa (possible non-specific binding), 90 kDa (possible non-specific binding)
Exposure time: 2 minutes
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Dot Blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)This image is courtesy of Christine Buerki and Karin Rothgiesser, University of Zurich, SwitzerlandRabbit polyclonal to NF-kB p65 (acetyl K310) (ab19870; 2.5µg/ml) in 1% non-fat milk TBS-T incubated for 3h at room temperature. Exposure time: 75 min normal ECL. This Dot blot demonstrates that ab19870 recognized upto 10ng of purified peptide on a PVDF membrane.
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Immunoprecipitation - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)This image is courtesy of Christine Buerki and Karin Rothgiesser, University of Zurich, SwitzerlandWestern Blot with ab19870 after p65 Immunoprecipitation: rabbit polyclonal to NF-kB p65 (acetyl K310) (ab19870; 2.5µg/ml) in 1% non-fat milk TBS-T incubated for 3 hours at room temperature. Exposure time: 1 min normal ECL. Tested samples: nuclear extracts (180 µg) of immortalized p65-/- mouse cells, complemented with the empty vector (pRRL), wild-type p65 (Wt) and non-acetylatable K310 (K310R). The samples tested were treated with deacetylase inhibitors HDACi (TSA + Nicotinamide) and TNF-alpha. The samples were immunoprecipitated with 2µg of alpha-p65 and subsequently analysed by Western blot with Rabbit polyclonal to NF-kB p65 (acetyl K310) (ab19870). Predicted band size = 65kDa, Observed band size = 75kDa. The p65 band runs higher in this SDS-PAGE blot as it contains a myc-tag.
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Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)This image is courtesy of Christine Buerki and Karin Rothgiesser, University of Zurich, SwitzerlandAll lanes : Anti-NF-kB p65 (acetyl K310) antibody (ab19870) at 2.5 µg/ml
Lane 1 : pRRL untreated
Lane 2 : pRRL HDACi
Lane 3 : pRRL HDACi + TNF
Lane 4 : Wt untreated
Lane 5 : Wt HDACi
Lane 6 : Wt HDACi + phorbol myristate acetate
Lane 7 : K310R untreated
Lane 8 : K310R HDACi
Lane 9 : K310R HDACi + phorbol myristate acetate
Lysates/proteins at 75 µg per lane.
Developed using the ECL technique.
Predicted band size: 65 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 1 hourab19870 recognizes Rabbit polyclonal to NF-kB p65 (acetyl K310) specifically at ~75kDa (indicated by the arrow) is this SDS-PAGE blot. The p65 band runs higher than 65kDa in this SDS-PAGE blot as it contains a myc-tag. We are sure that the band at ~75kDa is p65 since p65 specific antibodies detect the same band in IP and WB and there is no signal in the p65 knock-out cell line with ab19870. A number of additonal bands are recognized by ab19870 when tested with endogenous p65 from whole cell extracts, we do not know the identity of these bands.
Tested samples: nuclear extracts (75µg) of immortalized p65-/- mouse cells, complemented with the empty ve
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Western blot - Anti-NF-kB p65 (acetyl K310) antibody (ab19870)Anti-NF-kB p65 (acetyl K310) antibody (ab19870) at 1 µg/ml + Lung (Rat) Tissue Lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 65 kDa
Observed band size: 72 kDa why is the actual band size different from the predicted?
Additional bands at: 15 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 4 minutes
Protocols
Datasheets and documents
References
ab19870 has been referenced in 60 publications.
- Sanajou D et al. Reduction of renal tubular injury with a RAGE inhibitor FPS-ZM1, valsartan and their combination in streptozotocin-induced diabetes in the rat. Eur J Pharmacol 842:40-48 (2019). PubMed: 30393200
- Wang X et al. Astragaloside IV inhibits glucose-induced epithelial-mesenchymal transition of podocytes through autophagy enhancement via the SIRT-NF-?B p65 axis. Sci Rep 9:323 (2019). PubMed: 30674969
- Tan S et al. Curculigoside exerts significant anti-arthritic effects in vivo and in vitro via regulation of the JAK/STAT/NF-?B signaling pathway. Mol Med Rep 19:2057-2064 (2019). PubMed: 30664158
- Li J et al. Mechanisms of Aerobic Exercise Upregulating the Expression of Hippocampal Synaptic Plasticity-Associated Proteins in Diabetic Rats. Neural Plast 2019:7920540 (2019). PubMed: 30911292
- Liu HW et al. Exercise training upregulates SIRT1 to attenuate inflammation and metabolic dysfunction in kidney and liver of diabetic db/db mice. Nutr Metab (Lond) 16:22 (2019). PubMed: 30988688
