重组Anti-IKK beta抗体[Y466] (ab32135)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y466] to IKK beta
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-IKK beta抗体[Y466]
参阅全部 IKK beta 一抗 -
描述
兔单克隆抗体[Y466] to IKK beta -
宿主
Rabbit -
经测试应用
适用于: WB, IPmore details -
种属反应性
与反应: Mouse, Human -
免疫原
Synthetic peptide within Human IKK beta aa 650-750 (C terminal). The exact sequence is proprietary.
Database link: O14920 -
阳性对照
- WB: Wild-type HAP1 cell lysate. HeLa, NIH/3T3 and Daudi cell lysate. IHC-P: Human cervical carcinoma tissue. IP: SH-SY5Y whole cell lysate.
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常规说明
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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
Y466 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32135于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB | (3) |
1/1000 - 1/5000. Detects a band of approximately 87 kDa.
This antibody has weaker binding with mouse than human. |
IP |
1/50.
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说明 |
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WB
1/1000 - 1/5000. Detects a band of approximately 87 kDa. This antibody has weaker binding with mouse than human. |
IP
1/50. |
靶标
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功能
Acts as part of the IKK complex in the conventional pathway of NF-kappa-B activation and phosphorylates inhibitors of NF-kappa-B thus leading to the dissociation of the inhibitor/NF-kappa-B complex and ultimately the degradation of the inhibitor. Also phosphorylates NCOA3. -
组织特异性
Highly expressed in heart, placenta, skeletal muscle, kidney, pancreas, spleen, thymus, prostate, testis and peripheral blood. -
序列相似性
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. I-kappa-B kinase subfamily.
Contains 1 protein kinase domain. -
翻译后修饰
Upon cytokine stimulation, phosphorylated on Ser-177 and Ser-181 by MEKK1 and/or MAP3K14/NIK; which enhances activity. Once activated, autophosphorylates on the C-terminal serine cluster; which decreases activity and prevents prolonged activation of the inflammatory response.
Acetylation of Thr-180 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the I-kappa-B pathway.
Ubiquitinated. Monoubiquitination involves TRIM21 that leads to inhibition of Tax-induced NF-kappa-B signaling. According to PubMed:19675099, 'Ser-163' does not serve as a monoubiquitination site. According to PubMed:16267042, ubiquitination on 'Ser-163' modulates phosphorylation on C-terminal serine residues. Monoubiquitination by TRIM21 is dirupted by Yersinia yopJ. -
细胞定位
Cytoplasm. Membrane raft. Colocalized with DPP4 in membrane rafts. - Information by UniProt
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数据库链接
- Entrez Gene: 3551 Human
- Entrez Gene: 16150 Mouse
- Omim: 603258 Human
- SwissProt: O14920 Human
- SwissProt: O88351 Mouse
- Unigene: 597664 Human
- Unigene: 277886 Mouse
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别名
- I kappa B kinase 2 antibody
- I kappa B kinase beta antibody
- I-kappa-B kinase 2 antibody
see all
图片
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All lanes : Anti-IKK beta antibody [Y466] (ab32135) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : IKBKB CRISPR/Cas9 edited HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 87 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab32135 observed at 87 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32135 was shown to react with IKK beta in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab264847 (CRISPR/Cas9 edited cell lysate ab257228) lane below 87kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and IKBKB CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32135 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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ab32135 (purified) at 1:50 dilution (2μg) immunoprecipitating IKK beta in SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate.
Lane 1 (input): SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10ug
Lane 2 (+): ab32135 & SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32135 in SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysateFor western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:10,000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
Lanes 2, 4 and 6: IKK beta knockout HAP1 cell lysate (20 µg)
Lanes 1 and 2: Green signal from target – ab32135 observed at 87 kDa
Lanes 3 and 4: Red signal from loading control – ab8226 observed at 42 kDa
Lanes 5 and 6: Merged (red and green) signalUnpurified ab32135 was shown to react with IKK beta when IKK beta knockout samples were used, along with additional cross-reactive bands. Wild-type and IKK beta knockout samples were subjected to SDS-PAGE. ab32135 and ab8226 (loading control to beta actin) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4ºC. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
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All lanes : Anti-IKK beta antibody [Y466] (ab32135) at 1/1000 dilution (purified)
Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
Lane 2 : Mouse lung lysates
Lane 3 : Rat brain lysates
Lane 4 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 87 kDa why is the actual band size different from the predicted?Blocking and diluting buffer: 5% NFDM/TBST
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Anti-IKK beta antibody [Y466] (ab32135) at 1/2500 dilution (purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilutionBlocking and diluting buffer: 5% NFDM/TBST
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Anti-IKK beta antibody [Y466] (ab32135) at 1/5000 dilution (unpurified) + Daudi cell lysate
Observed band size: 87 kDa why is the actual band size different from the predicted? -
unpurified ab32135
实验方案
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.
数据表及文件
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SDS download
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Datasheet download
文献 (49)
ab32135 被引用在 49 文献中.
- Su Y et al. Effects of Fish Oil, Lipid Mediators, Derived from Docosahexaenoic Acid, and Their Co-Treatment against Lipid Metabolism Dysfunction and Inflammation in HFD Mice and HepG2 Cells. Nutrients 15:N/A (2023). PubMed: 36678298
- Cariulo C et al. IKBKB reduces huntingtin aggregation by phosphorylating serine 13 via a non-canonical IKK pathway. Life Sci Alliance 6:N/A (2023). PubMed: 37553253
- Tian Y et al. Three Chinese pedigrees of A20 haploinsufficiency: clinical, cytokine and molecular characterization. Front Immunol 13:955079 (2022). PubMed: 35958611
- Guo Y et al. Tripartite motif 52 (TRIM52) promotes proliferation, migration, and regulation of colon cancer cells associated with the NF-κB signaling pathway. J Gastrointest Oncol 13:1097-1111 (2022). PubMed: 35837156
- Zhang R et al. The Endoplasmic Reticulum ATP13A1 is Essential for MAVS-Mediated Antiviral Innate Immunity. Adv Sci (Weinh) 9:e2203831 (2022). PubMed: 36216581