重组Anti-PHD3抗体[EPR17869] (ab184714)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17869] to PHD3
- Suitable for: WB, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-PHD3抗体[EPR17869]
参阅全部 PHD3 一抗 -
描述
兔单克隆抗体[EPR17869] to PHD3 -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IF, IPmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: PHD3 transfected HEK-293 whole cell lysate and PHD3 transfected HEK-293 whole cell lysate treated with 0.1 mM CoCl2 (Cobalt (II) chloride) for 4 hours. Human fetal liver lysate. A549, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates. Mouse pancreas, kidney and spleen lysates and Rat pancreas and brain lysates. MCF7 cell lysate treated with 0.5mM CoCl2 (Cobalt (II) chloride) for 6 hours. ICC/IF: A549 and PC-12 cells. IP: NIH/3T3 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 (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR17869 -
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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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab184714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 27 kDa (predicted molecular weight: 27 kDa).
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ICC/IF |
1/250.
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IP |
1/70.
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说明 |
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WB
1/2000. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa). |
ICC/IF
1/250. |
IP
1/70. |
靶标
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功能
Catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates HIF-1 alpha at 'Pro-564', and HIF-2 alpha. Functions as a cellular oxygen sensor and, under normoxic conditions, targets HIF through the hydroxylation for proteasomal degradation via the von Hippel-Lindau ubiquitination complex. May play a role in cell growth regulation in muscle cells and in apoptosis in neuronal tissue. Promotes cell death through a caspase-dependent mechanism. -
组织特异性
Widely expressed at low levels. Expressed at higher levels in heart (cardiac myocytes, aortic endothelial cells and coronary artery smooth muscle) and placenta. -
序列相似性
Contains 1 Fe2OG dioxygenase domain. -
细胞定位
Cytoplasm. Nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 112399 Human
- Entrez Gene: 112407 Mouse
- Entrez Gene: 54702 Rat
- Omim: 606426 Human
- SwissProt: Q9H6Z9 Human
- SwissProt: Q91UZ4 Mouse
- SwissProt: Q62630 Rat
- Unigene: 135507 Human
see all -
别名
- Egl 9 family hypoxia inducible factor 3 antibody
- Egl nine homolog 3 (C. elegans) antibody
- Egl nine homolog 3 antibody
see all
图片
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All lanes : Anti-PHD3 antibody [EPR17869] (ab184714) at 1/10000 dilution
Lane 1 : PHD3 transfected HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
Lane 2 : Empty vector (vector control) transfected HEK-293 whole cell lysate
Lane 3 : PHD3 transfected HEK-293 whole cell lysate treated with 0.1 mM CoCl2 for 4 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-PHD3 antibody [EPR17869] (ab184714) at 1/2000 dilution + Human fetal liver lysate at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-PHD3 antibody [EPR17869] (ab184714) at 1/5000 dilution + A549 (Human lung carcinoma) whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 2 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-PHD3 antibody [EPR17869] (ab184714) at 1/2000 dilution
Lane 1 : Mouse pancreas lysate
Lane 2 : Rat pancreas lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 1 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-PHD3 antibody [EPR17869] (ab184714) at 1/2000 dilution
Lane 1 : Mouse kidney lysate
Lane 2 : Mouse spleen lysate
Lane 3 : Rat brain lysate
Lane 4 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 6 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-PHD3 antibody [EPR17869] (ab184714) at 1/2000 dilution
Lane 1 : Untreated MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 2 : MCF7 cell lysate treated with 0.5mM CoCl2 (Cobalt (II) chloride) for 6 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 27 kDa
Observed band size: 27 kDa
Exposure time: 1 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
PHD3 expression was induced by CoCl2 treatment (PMID: 18337469).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (Human lung carcinoma) cells labeling PHD3 with ab184714 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing weakly cytoplasm and nuclear staining on A549 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab184714 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-12 (Rat adrenal gland pheochromocytoma) cells labeling PHD3 with ab184714 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing weakly cytoplasm and nuclear staining on PC-12 cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab184714 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
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PHD3 was immunoprecipitated from 1mg of NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate with ab184714 at 1/70 dilution.
Western blot was performed from the immunoprecipitate using ab184714 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/10000 dilution.
Lane 1: NIH/3T3 whole cell lysate 10ug (Input).
Lane 2: ab184714 IP in NIH/3T3 whole cell lysate.
Lane 3: NIH/3T3 whole cell lysate supernatant after capture (unbound).
Lane 4: Rabbit monoclonal IgG (ab172730) instead of ab184714 in NIH/3T3 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
ab184714 is not a strong binder for IP - only a partial amount of the target protein in the lysate was immune-precipitated.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (4)
ab184714 被引用在 4 文献中.
- Liu Q et al. Insights into male androgenetic alopecia using comparative transcriptome profiling: hypoxia-inducible factor-1 and Wnt/β-catenin signalling pathways. Br J Dermatol 187:936-947 (2022). PubMed: 35862273
- Zacharias NM et al. Prolyl Hydroxylase 3 Knockdown Accelerates VHL-Mutant Kidney Cancer Growth In Vivo. Int J Mol Sci 22:N/A (2021). PubMed: 33799686
- Fan S et al. TET is targeted for proteasomal degradation by the PHD-pVHL pathway to reduce DNA hydroxymethylation. J Biol Chem 295:16299-16313 (2020). PubMed: 32963106
- Tavernier SJ et al. Opposing regulation and roles for PHD3 in lung dendritic cells and alveolar macrophages. J Leukoc Biol 102:1115-1126 (2017). PubMed: 28716863