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
- 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 blotmore details
Unsuitable for: ICC/IF -
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. |
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
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
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
