重组Anti-PPP2R5D抗体[EPR15617] (ab188323)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR15617] to PPP2R5D
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Knockout validated
- Reacts with: Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-PPP2R5D抗体[EPR15617]
参阅全部 PPP2R5D 一抗 -
描述
兔单克隆抗体[EPR15617] to PPP2R5D -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
种属反应性
与反应: Rat, Human
预测可用于: Mouse -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: A431, MCF7, C6, HepG2, Jurkat, HeLa, 293T cell lysate. IHC-P: Human brain tissue. IP: HEK293 cells. Flow Cyt (intra): HeLa cells.
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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
EPR15617 -
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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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab188323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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Flow Cyt (Intra) |
1/40.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/10000 - 1/50000. Detects a band of approximately 58, 66, 70 kDa (predicted molecular weight: 70 kDa).
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/250.
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IP |
1/40 - 1/60.
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说明 |
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Flow Cyt (Intra)
1/40. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/10000 - 1/50000. Detects a band of approximately 58, 66, 70 kDa (predicted molecular weight: 70 kDa). |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/250. |
IP
1/40 - 1/60. |
靶标
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功能
The B regulatory subunit might modulate substrate selectivity and catalytic activity, and also might direct the localization of the catalytic enzyme to a particular subcellular compartment. -
组织特异性
Isoform Delta-2 is widely expressed. Isoform Delta-1 is highly expressed in brain. -
序列相似性
Belongs to the phosphatase 2A regulatory subunit B56 family. -
细胞定位
Cytoplasm. Nucleus. Nuclear in interphase, nuclear during mitosis. - Information by UniProt
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数据库链接
- Entrez Gene: 5528 Human
- Entrez Gene: 21770 Mouse
- Entrez Gene: 363193 Rat
- Omim: 601646 Human
- SwissProt: Q14738 Human
- Unigene: 533308 Human
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别名
- 2A5D_HUMAN antibody
- B'delta antibody
- B56D antibody
see all
图片
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All lanes : Anti-PPP2R5D antibody [EPR15617] (ab188323) at 1/10000 dilution
Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : PPP2R5D knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 70 kDa
Observed band size: 60-65 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab188323 observed at 60-65 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab188323 was shown to react with PPP2R5D in wild-type A431 cells in western blot with loss of signal observed in PPP2R5D knockout sample. Wild-type and PPP2R5D knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab188323 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemical analysis of paraffin-embedded Human brain tissue labeling PPP2R5D using ab188323 at 1/100 dilution. A Ready to use HRP Polymer for Rabbit IgG (prediluted) was used as secondary. Counterstain: Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-PPP2R5D antibody [EPR15617] (ab188323) at 1/100000 dilution
Lane 1 : C6 cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : 293T cell lysate
Lane 4 : HeLa cell lysate
Lane 5 : Jurkat 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: 70 kDa
Observed band size: 70 kDaBased on the sequence analysis, ab188323 recognizes 3 isoforms with the predicted MWs of 58KDa, 66KDa and 70KDa, respectively.
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Immunoprecipitation analysis of 293T cell lysate labeling PPP2R5D using ab188323 at 1/50 dilution. A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1500 was used as secondary antibody.
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Immunofluorescence analysis of HepG2 cells labeling PPP2R5D using ab188323 at 1/250 dilution. A Goat anti rabbit IgG (Alexa Fluor488) at 1/200 dilution was used as secondary antibody. Cells were fixed with 4% Paraformaldehyde. Counterstain: DAPI.
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Intracellular Flow Cytometry analysis of HeLa cells labeling PPP2R5D using ab188323 at 1/40 dilution (pink). A Goat anti rabbit IgG (FITC) at 1/150 dilution was used as secondary antibody. Cells were fixed with 2% paraformaldehyde. Isotype control: Rabbit monoclonal IgG (green).
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 (6)
ab188323 被引用在 6 文献中.
- Papke CM et al. A disorder-related variant (E420K) of a PP2A-regulatory subunit (PPP2R5D) causes constitutively active AKT-mTOR signaling and uncoordinated cell growth. J Biol Chem 296:100313 (2021). PubMed: 33482199
- Jong CJ et al. Reduction of protein phosphatase 2A (PP2A) complexity reveals cellular functions and dephosphorylation motifs of the PP2A/B'd holoenzyme. J Biol Chem 295:5654-5668 (2020). PubMed: 32156701
- Marelli S et al. Antagonism of PP2A is an independent and conserved function of HIV-1 Vif and causes cell cycle arrest. Elife 9:N/A (2020). PubMed: 32292164
- Naamati A et al. Functional proteomic atlas of HIV infection in primary human CD4+ T cells. Elife 8:N/A (2019). PubMed: 30857592
- Evans EL et al. HIV-1 Vif's Capacity To Manipulate the Cell Cycle Is Species Specific. J Virol 92:N/A (2018). PubMed: 29321323
- Greenwood EJ et al. Temporal proteomic analysis of HIV infection reveals remodelling of the host phosphoproteome by lentiviral Vif variants. Elife 5:N/A (2016). PubMed: 27690223