Anti-NF-kB p65 (phospho S536) 抗体 [EP2294Y] (ab76302)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2294Y] to NF-kB p65 (phospho S536)
- Suitable for: Dot blot, WB, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
-
製品名
Anti-NF-kB p65 (phospho S536) antibody [EP2294Y]
NF-kB p65 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP2294Y] to NF-kB p65 (phospho S536) -
由来種
Rabbit -
特異性
Stimulation may be required to allow detection of the phosphorylated protein.
We recommend using NIH/3T3 (Mouse embryonic fibroblast) treated with 100nM Calyculin A for 30 minutes as a positive control.
-
アプリケーション
適用あり: Dot blot, WB, IPmore details
適用なし: Flow Cyt,ICC/IF or IHC-P -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
ポジティブ・コントロール
- WB: HeLa whole cell lysate treated with Calyculin A + TNF-alpha. C6 and NIH/3T3 treated with 100nM Calyculin A for 30 minutes whole cell lysate . IP: Daudi cell lysate treated with Calyculin A + TNF-alpha.
-
特記事項
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
-
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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP2294Y -
アイソタイプ
IgG -
研究分野
関連製品
-
Alternative Versions
-
Assay kits
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
-
Recombinant Protein
-
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ab76302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 |
---|---|---|
Dot blot |
1/1000.
|
|
WB | (7) |
1/1000. Predicted molecular weight: 65 kDa.
|
IP |
1/20 - 1/30.
|
特記事項 |
---|
Dot blot
1/1000. |
WB
1/1000. Predicted molecular weight: 65 kDa. |
IP
1/20 - 1/30. |
ターゲット情報
-
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
-
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
-
別名
- Avian reticuloendotheliosis viral (v rel) oncogene homolog A antibody
- MGC131774 antibody
- NF kappa B p65delta3 antibody
see all
画像
-
ab76302 at 1/30 immunoprecipitating NF-kB p65 (phospho S536) in Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha.
Lane 1 (input): Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha (10µg)
Lane 2 (+): ab76302 + Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76302 in Daudi whole cell lysate treated with calyculin A and tumor necrosis factor-alpha.
For western blotting, ab76302 at 1/500 dilution (0.95 μg/ml) and VeriBlot for IP Detection Reagent (HRP)(ab131366) at 1/1000 dilution were used.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
-
All lanes : Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] (ab76302) at 1/1000 dilution
Lane 1 : C6 (Rat glial tumor glial cell) whole cell lysate
Lane 2 : C6 (Rat glial tumor glial cell) treated with 100nM Calyculin A for 30 minutes whole cell lysate
Lane 3 : C6 (Rat glial tumor glial cell) treated with 100nM Calyculin A for 30 minutes, then the membrane treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 65 kDa
Observed band size: 65 kDa -
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 100nM Calyculin A for 30 minutes whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) treated with 100nM Calyculin A for 30 minutes, then the membrane treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 65 kDa
Observed band size: 65 kDa -
All lanes : Anti-NF-kB p65 (phospho S536) antibody [EP2294Y] (ab76302) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with Calyculin A and TNF-alpha whole cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 65 kDa
Observed band size: 65 kDa -
Dot blot analysis of INF- kB p65 (phospho S536) phospho peptide (Lane 1) and NF- kB p65 non-phospho peptide (Lane 2) labeling NF-kB p65 (phospho S536) with ab76302 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG) (H+L) at 1/100 000 was used as the secondary antibody.
Blocking and diluting buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
プロトコール
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (160)
ab76302 は 160 報の論文で使用されています。
- Lin H et al. Low-intensity pulsed ultrasound enhances immunomodulation and facilitates osteogenesis of human periodontal ligament stem cells by inhibiting the NF-κB pathway. Cell Tissue Bank 24:45-58 (2023). PubMed: 35644018
- Wei Z et al. Inhibition of secretory leukocyte protease inhibitor (SLPI) promotes the PUMA-mediated apoptosis and chemosensitivity to cisplatin in colorectal cancer cells. Discov Oncol 14:1 (2023). PubMed: 36595102
- Wu Q et al. UBR5 promotes migration and invasion of glioma cells by regulating the ECRG4/NF-ҡB pathway. J Biosci 47:N/A (2022). WB ; Human . PubMed: 36226349
- Li J et al. METTL3 promotes prostatic hyperplasia by regulating PTEN expression in an m6A-YTHDF2-dependent manner. Cell Death Dis 13:723 (2022). WB ; Human . PubMed: 35985997
- Ou D et al. LIM mineralization protein-1 inhibits IL-1β-induced human chondrocytes injury by altering the NF-κB and MAPK/JNK pathways. Exp Ther Med 23:61 (2022). PubMed: 34934432