Anti-NF-kB p65 (acetyl K310) 抗体 (ab19870)
Key features and details
- Rabbit polyclonal to NF-kB p65 (acetyl K310)
- Suitable for: WB, IP, Dot blot
- Reacts with: Mouse, Rat
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
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製品名
Anti-NF-kB p65 (acetyl K310) antibody
NF-kB p65 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to NF-kB p65 (acetyl K310) -
由来種
Rabbit -
アプリケーション
適用あり: WB, IP, Dot blotmore details
適用なし: ICC/IF -
種交差性
交差種: Mouse, Rat
交差が予測される動物種: Human
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免疫原
Synthetic peptide corresponding to Human NF-kB p65 aa 300-400 (internal sequence) conjugated to keyhole limpet haemocyanin.
(Peptide available asab20612) -
特記事項
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
バッファー
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... -
精製度
Protein A purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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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
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab19870の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| WB | (3) |
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)
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| IP |
Use a concentration of 2.5 µg/ml.
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| Dot blot |
Use at an assay dependent concentration.
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| 特記事項 |
|---|
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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) |
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IP
Use a concentration of 2.5 µg/ml. |
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Dot blot
Use at an assay dependent concentration. |
ターゲット情報
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機能
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. -
配列類似性
Contains 1 RHD (Rel-like) domain. -
ドメイン
the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors. -
翻訳後修飾
Ubiquitinated, 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'. -
細胞内局在
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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参照データベース
- 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
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別名
- Avian reticuloendotheliosis viral (v rel) oncogene homolog A antibody
- MGC131774 antibody
- NF kappa B p65delta3 antibody
see all
画像
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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
プロトコール
データシートおよび資料
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Datasheet download
参考文献 (92)
ab19870 は 92 報の論文で使用されています。
- Ma Q et al. MicroRNA-210 downregulates TET2 and contributes to inflammatory response in neonatal hypoxic-ischemic brain injury. J Neuroinflammation 18:6 (2021). PubMed: 33402183
- Lee SU et al. Transforming Growth Factor ß Inhibits MUC5AC Expression by Smad3/HDAC2 Complex Formation and NF-?B Deacetylation at K310 in NCI-H292 Cells. Mol Cells 44:38-49 (2021). PubMed: 33510050
- Chen L et al. Mangiferin prevents myocardial infarction-induced apoptosis and heart failure in mice by activating the Sirt1/FoxO3a pathway. J Cell Mol Med 25:2944-2955 (2021). PubMed: 33523605
- Zhou B et al. Bortezomib suppresses self-renewal and leukemogenesis of leukemia stem cell by NF-?B-dependent inhibition of CDK6 in MLL-rearranged myeloid leukemia. J Cell Mol Med 25:3124-3135 (2021). PubMed: 33599085
- Cheng Y et al. iTRAQ-Based Quantitative Proteomics Indicated Nrf2/OPTN-Mediated Mitophagy Inhibits NLRP3 Inflammasome Activation after Intracerebral Hemorrhage. Oxid Med Cell Longev 2021:6630281 (2021). PubMed: 33628368
