重组Anti-PKD2抗体[EP1495Y] (ab51250)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1495Y] to PKD2
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-PKD2抗体[EP1495Y]
参阅全部 PKD2 一抗 -
描述
兔单克隆抗体[EP1495Y] to PKD2 -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, ICC/IFmore details
不适用于: IP -
种属反应性
与反应: Mouse, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- HeLa whole cell lysate (ab150035) or human breast carcinoma tissue. IF/ICC: HeLa cell line
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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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% 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
EP1495Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Isotype control
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Positive Controls
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab51250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 | (10) |
1/500. Detects a band of approximately 105 kDa (predicted molecular weight: 97 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF | (2) |
1/100 - 1/250.
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说明 |
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WB
1/500. Detects a band of approximately 105 kDa (predicted molecular weight: 97 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/250. |
靶标
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功能
Converts transient diacylglycerol (DAG) signals into prolonged physiological effects, downstream of PKC. Involved in resistance to oxidative stress. -
组织特异性
Widely expressed. -
序列相似性
Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. PKD subfamily.
Contains 1 PH domain.
Contains 2 phorbol-ester/DAG-type zinc fingers.
Contains 1 protein kinase domain. -
翻译后修饰
Phosphorylation of Ser-876 correlates with the activation status of the kinase.
Ser-706 is probably phosphorylated by PKC. -
细胞定位
Cytoplasm. Membrane. Translocation to the cell membrane is required for kinase activation. - Information by UniProt
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数据库链接
- Entrez Gene: 25865 Human
- Entrez Gene: 101540 Mouse
- Omim: 607074 Human
- SwissProt: Q9BZL6 Human
- SwissProt: Q8BZ03 Mouse
- Unigene: 466987 Human
- Unigene: 1881 Mouse
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别名
- DKFZp586E0820 antibody
- HSPC187 antibody
- KPCD2_HUMAN antibody
see all
图片
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All lanes : Anti-PKD2 antibody [EP1495Y] (ab51250) at 1/500 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : PRKD2 knockout A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 97 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?Western blot: Anti-PRKD2 antibody [EP1495Y] (ab51250) staining at 1/500 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab51250 was shown to bind specifically to PRKD2. A band was observed at 105 kDa in wild-type A549 cell lysates with no signal observed at this size in PRKD2 knockout cell line. To generate this image, wild-type and PRKD2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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Lane 1: Wild-type HAP1 cell lysate (40 µg)
Lane 2: PKD2 knockout HAP1 cell lysate (40 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: HEK293 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab51250 observed at 110 kDa. Red - loading control, ab8245, observed at 37 kDa.ab51250 was shown to recognize PKD2 when PKD2 knockout samples were used, along with additional cross-reactive bands. Wild-type and PKD2 knockout samples were subjected to SDS-PAGE. Ab51250 and ab8245 (loading control to GAPDH) were diluted at 1/500 and 1:10,000 dilution respectively and incubated overnight at 4C. 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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Anti-PKD2 antibody [EP1495Y] (ab51250) at 1/500 dilution + HeLa cell lysate at 10 µg
Secondary
Goat anti-rabbit HRP labeled at 1/2000 dilution
Predicted band size: 97 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKD2 antibody [EP1495Y] (ab51250)
Ab51250 (1:100) staining human PKD2 in human breast carcinoma tissue by immunohistochemistry using paraffin embedded tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF image of ab51250 stained HeLa cells. The cells were 4% Formaldehyde fixed (10 mins) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51250, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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 (8)
ab51250 被引用在 8 文献中.
- Leightner AC et al. Regulation of Osteoclast Differentiation at Multiple Stages by Protein Kinase D Family Kinases. Int J Mol Sci 21:N/A (2020). PubMed: 32033440
- Zhu Z et al. Inhibition of protein kinase D by CID755673 promotes maintenance of the pluripotency of embryonic stem cells. Development 147:N/A (2020). PubMed: 32747433
- Baker J et al. Protein kinase D3 modulates MMP1 and MMP13 expression in human chondrocytes. PLoS One 13:e0195864 (2018). PubMed: 29652915
- Plastira I et al. Lysophosphatidic acid via LPA-receptor 5/protein kinase D-dependent pathways induces a motile and pro-inflammatory microglial phenotype. J Neuroinflammation 14:253 (2017). PubMed: 29258556
- Azoitei N et al. HSP90 supports tumor growth and angiogenesis through PRKD2 protein stabilization. Cancer Res 74:7125-36 (2014). PubMed: 25297628
- Bhavanasi D et al. Protein kinase Cd mediates the activation of protein kinase D2 in platelets. Biochem Pharmacol 82:720-7 (2011). WB ; Human, Mouse . PubMed: 21736870
- Azoitei N et al. Protein kinase D2 is a novel regulator of glioblastoma growth and tumor formation. Neuro Oncol 13:710-24 (2011). WB, IHC-P ; Human, Chicken . PubMed: 21727210
- Azoitei N et al. Protein kinase D2 is a crucial regulator of tumour cell-endothelial cell communication in gastrointestinal tumours. Gut 59:1316-30 (2010). PubMed: 20732914