Anti-IKB alpha 抗体 [EP697] (ab76429)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP697] to IKB alpha
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-IKB alpha antibody [EP697]
IKB alpha 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP697] to IKB alpha -
由来種
Rabbit -
アプリケーション
適用あり: WB, IHC-P, ICC/IFmore details
適用なし: Flow Cyt -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human IKB alpha (C terminal). The exact sequence is proprietary.
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ポジティブ・コントロール
- HeLa, PC-12, RAW264.7 and Jurkat cell lysates; human kidney and colon tissue.
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特記事項
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.
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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP697 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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ab76429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 |
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WB | (1) |
1/500 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
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IHC-P |
1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
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ICC/IF |
1/250.
For unpurified use at 1/50 - 1/100. |
特記事項 |
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WB
1/500 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa). |
IHC-P
1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
ICC/IF
1/250. For unpurified use at 1/50 - 1/100. |
ターゲット情報
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機能
Inhibits the activity of dimeric NF-kappa-B/REL complexes by trapping REL dimers in the cytoplasm through masking of their nuclear localization signals. On cellular stimulation by immune and proinflammatory responses, becomes phosphorylated promoting ubiquitination and degradation, enabling the dimeric RELA to translocate to the nucleus and activate transcription. -
関連疾患
Ectodermal dysplasia, anhidrotic, with T-cell immunodeficiency autosomal dominant -
配列類似性
Belongs to the NF-kappa-B inhibitor family.
Contains 5 ANK repeats. -
翻訳後修飾
Phosphorylated; disables inhibition of NF-kappa-B DNA-binding activity. Phosphorylation at positions 32 and 36 is prerequisite to recognition by UBE2D3 leading to polyubiquitination and subsequent degradation.
Sumoylated; sumoylation requires the presence of the nuclear import signal. Sumoylation blocks ubiquitination and proteasome-mediated degradation of the protein thereby increasing the protein stability.
Monoubiquitinated at Lys-21 and/or Lys-22 by UBE2D3. Ubiquitin chain elongation is then performed by CDC34 in cooperation with the SCF(FBXW11) E3 ligase complex, building ubiquitin chains from the UBE2D3-primed NFKBIA-linked ubiquitin. The resulting polyubiquitination leads to protein degradation. Also ubiquitinated by SCF(BTRC) following stimulus-dependent phosphorylation at Ser-32 and Ser-36.
Deubiquitinated by porcine reproductive and respiratory syndrome virus Nsp2 protein, which thereby interferes with NFKBIA degradation and impairs subsequent NF-kappa-B activation. -
細胞内局在
Cytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm by a nuclear localization signal (NLS) and a CRM1-dependent nuclear export. - Information by UniProt
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参照データベース
- Entrez Gene: 4792 Human
- Entrez Gene: 18035 Mouse
- Entrez Gene: 25493 Rat
- Omim: 164008 Human
- SwissProt: P25963 Human
- SwissProt: Q9Z1E3 Mouse
- SwissProt: Q63746 Rat
- Unigene: 81328 Human
see all -
別名
- I kappa B alpha antibody
- I-kappa-B-alpha antibody
- IkappaBalpha antibody
see all
画像
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: IKB alpha knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)Lanes 1 - 3: Merged signal (red and green). Green - ab76429 observed at 39 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab76429 was shown to specifically react with IKB alpha in wild-type HAP1 cells as well as additional cross-reactive bands. No bands were observed when IKB alpha knockout cells were examined. Wild-type and IKB alpha knockout samples were subjected to SDS-PAGE. Ab76429 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/500 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-IKB alpha antibody [EP697] (ab76429) at 1/1000 dilution
Lane 1 : Jurkat (human acute T cell leukemia) whole cell lysate
Lane 2 : HeLa (human cervix adenocarcinoma) 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: 36 kDa
Observed band size: 39 kDa why is the actual band size different from the predicted?
Blocking/Diluting buffer 5% NFDM /TBST -
Immunohistochemical analysis of paraffin-embedded human colon tissue sections labelling IKB alpha with purified ab76429 at dilution of 1/500. Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
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Immunocytochemistry/Immunofluorescence staining of HeLa cells labelling IKB alpha with purified ab76429 at a working dilution of 1/250. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. DAPI was used as nuclear counterstain. The cells were fixed in 4% Paraformaldehyde and permeabilized using 0.1% Triton X-100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, rabbit primary antibody was used followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120). For negative control 2, ab7291 (mouse anti-tubulin) was used followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077).
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All lanes : Anti-IKB alpha antibody [EP697] (ab76429) at 1/10000 dilution
Lane 1 : RAW264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate
Lane 2 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 36 kDa
Observed band size: 39 kDa why is the actual band size different from the predicted?
Blocking/Diluting buffer 5% NFDM /TBST -
Immunohistochemical analysis of Paraffin-embedded human colon tissue section labelling IKB alpha with ab76429 at 1/100 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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All lanes : Anti-IKB alpha antibody [EP697] (ab76429) at 1/1000 dilution
Lane 1 : HeLa cell lysates
Lane 2 : Jurkat cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : goat anti-rabbit HRP at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (23)
ab76429 は 23 報の論文で使用されています。
- Li D et al. PFKFB4 promotes angiogenesis via IL-6/STAT5A/P-STAT5 signaling in breast cancer. J Cancer 13:212-224 (2022). PubMed: 34976184
- Shi L et al. Qingwei San treats oral ulcer subjected to stomach heat syndrome in db/db mice by targeting TLR4/MyD88/NF-κB pathway. Chin Med 17:1 (2022). PubMed: 34980192
- Duan Y et al. Mulberry Leaf Flavonoids Inhibit Liver Inflammation in Type 2 Diabetes Rats by Regulating TLR4/MyD88/NF-κB Signaling Pathway. Evid Based Complement Alternat Med 2022:3354062 (2022). PubMed: 35845591
- Shen H et al. TRIM46 aggravated high glucose-induced hyper permeability and inflammatory response in human retinal capillary endothelial cells by promoting IκBα ubiquitination. Eye Vis (Lond) 9:35 (2022). PubMed: 36064447
- Chen J et al. Regulation of Atherosclerosis by Toll-Like Receptor 4 Induced by Serum Amyloid 1: A Systematic In Vitro Study. Biomed Res Int 2022:4887593 (2022). PubMed: 36158875