重组Anti-PDK1抗体[EPR19573] (ab207450)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19573] to PDK1
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-PDK1抗体[EPR19573]
参阅全部 PDK1 一抗 -
描述
兔单克隆抗体[EPR19573] to PDK1 -
宿主
Rabbit -
经测试应用
适用于: WB, IPmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa, Hap1, HEK-293, Jurkat, HepG2, NIH/3T3, PC-12 and LNCaP whole cell lysates; human heart and kidney lysates; mouse brain, heart, kidney and spleen lysates; rat kidney and spleen lysates; HEK-293 mitochondria fraction lysate. IP: HepG2 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.
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 long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% 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
EPR19573 -
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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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab207450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 |
1/2000. Detects a band of approximately 47 kDa (predicted molecular weight: 49 kDa).
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IP |
1/40.
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说明 |
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WB
1/2000. Detects a band of approximately 47 kDa (predicted molecular weight: 49 kDa). |
IP
1/40. |
靶标
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功能
Inhibits the mitochondrial pyruvate dehydrogenase complex by phosphorylation of the E1 alpha subunit, thus contributing to the regulation of glucose metabolism. -
组织特异性
Expressed predominantly in the heart. -
序列相似性
Belongs to the PDK/BCKDK protein kinase family.
Contains 1 histidine kinase domain. -
细胞定位
Mitochondrion matrix. - Information by UniProt
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数据库链接
- Entrez Gene: 5163 Human
- Entrez Gene: 228026 Mouse
- Entrez Gene: 116551 Rat
- Omim: 602524 Human
- SwissProt: Q15118 Human
- SwissProt: Q8BFP9 Mouse
- SwissProt: Q63065 Rat
- Unigene: 470633 Human
see all -
别名
- [Pyruvate dehydrogenase [lipoamide]] kinase isozyme 1, mitochondrial antibody
- HGNC:8809 antibody
- Mitochondrial pyruvate dehydrogenase kinase isoenzyme 1 antibody
see all
图片
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All lanes : Anti-PDK1 antibody [EPR19573] (ab207450) at 1/2000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PDK1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 49 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab207450 observed at 45 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab207450 was shown to react with PDK1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264791 (knockout cell lysate ab257164) was used. Wild-type HeLa and PDK1 knockout 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. ab207450 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 2000 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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All lanes : Anti-PDK1 antibody [EPR19573] (ab207450) at 1/2000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : PDK1 knockout HAP1 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : LnCaP cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 49 kDaLanes 1 - 4: Merged signal (red and green). Green - ab207450 observed at 45 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab207450 was shown to specifically react with PDK1 in wild-type HAP1 cells. No band was observed when PDK1 knockout samples were examined. Wild-type and PDK1 knockout samples were subjected to SDS-PAGE. Ab207450 and ab8245 (loading control to GAPDH) were diluted at 1/2000 and 1/10,000 dilution respectively and incubated overnight at 4°C. 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 hour at room temperature before imaging.
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PDK1 was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab207450 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab207450 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HepG2 whole cell lysate 10µg (Input).
Lane 2: ab207450 IP in HepG2 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207450 in HepG2 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
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All lanes : Anti-PDK1 antibody [EPR19573] (ab207450) at 1/2000 dilution
Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 6 : LNCaP (Human prostate cancer cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 49 kDa
Observed band size: 47 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1-5: 15 seconds; Lane 6: 8 seconds.
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All lanes : Anti-PDK1 antibody [EPR19573] (ab207450) at 1/2000 dilution
Lane 1 : Human heart lysate
Lane 2 : Human kidney lysate
Lane 3 : Mouse brain lysate
Lane 4 : Mouse heart lysate
Lane 5 : Mouse kidney lysate
Lane 6 : Mouse spleen lysate
Lane 7 : Rat kidney lysate
Lane 8 : Rat spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
Lanes 1-2 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Lanes 3-8 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 49 kDa
Observed band size: 47 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/3/4: 8 seconds; Lane 2: 30 seconds; Lane 5/6: 3 minutes; Lane 7/8: 15 seconds.
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All lanes : Anti-PDK1 antibody [EPR19573] (ab207450) at 1/5000 dilution
Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) mitochondria fraction lysate
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) cytosol fraction lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 49 kDa
Observed band size: 47 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (18)
ab207450 被引用在 18 文献中.
- Tang HY et al. PDGFRβ modulates aerobic glycolysis in osteosarcoma HOS cells via the PI3K/AKT/mTOR/c-Myc pathway. Biochem Cell Biol 100:75-84 (2022). PubMed: 34678088
- Wang R et al. TRIM35 functions as a novel tumor suppressor in breast cancer by inducing cell apoptosis through ubiquitination of PDK1. Neoplasma 69:370-382 (2022). PubMed: 35081724
- Guo JQ et al. PDGF-BB/PDGFRβ promotes epithelial-mesenchymal transition by affecting PI3K/AKT/mTOR-driven aerobic glycolysis in Wilms' tumor G401 cells. Cell Biol Int 46:907-921 (2022). PubMed: 35165984
- Liu Y et al. Pharmacological inhibition of sphingolipid synthesis reduces ferroptosis by stimulating the HIF-1 pathway. iScience 25:104533 (2022). PubMed: 35784791
- Larrieu CM et al. Refining the Role of Pyruvate Dehydrogenase Kinases in Glioblastoma Development. Cancers (Basel) 14:N/A (2022). PubMed: 35954433