重组Anti-Hamartin抗体[EP318Y] (ab40872)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP318Y] to Hamartin
- Suitable for: IP, WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
选择批间可重复性更高的重组抗体
- 研究可靠 —— 各批次间结果一致且可重复
- 长期批量供应 —— 采用重组技术,可实现快速生产
- 首次实验即可成功 —— 经过大量验证确认了特异性
- 符合伦理标准 —— 产品不含动物成分
概述
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产品名称
Anti-Hamartin抗体[EP318Y]
参阅全部 Hamartin 一抗 -
描述
兔单克隆抗体[EP318Y] to Hamartin -
宿主
Rabbit -
经测试应用
适用于: IP, WB, IHC-P, Flow Cyt (Intra)more details
不适用于: ICC/IF -
种属反应性
与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- IHC-P: human liver tissue WB: HeLa cells; wild type HAP1 cell lysate, Hamartin knockout HAP1 cell lysate, HeLa cells, Human skeletal muscle tissue lysate 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. 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: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP318Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 488 Anti-Hamartin antibody [EP318Y] (ab223387)
- Alexa Fluor® 647 Anti-Hamartin antibody [EP318Y] (ab223388)
- Anti-Hamartin antibody [EP318Y] - BSA and Azide free (ab247297)
- Alexa Fluor® 568 Anti-Hamartin antibody [EP318Y] (ab312426)
- Alexa Fluor® 750 Anti-Hamartin antibody [EP318Y] (ab321626)
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Compatible Secondaries
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab40872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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IP |
1/500.
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WB |
1/1000. Predicted molecular weight: 150 kDa.
For the unpurified version use 1/20000 dilution |
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IHC-P |
1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For the unpurified version use 1/1000- 1/10000 dilution |
说明 |
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IP
1/500. |
WB
1/1000. Predicted molecular weight: 150 kDa. For the unpurified version use 1/20000 dilution |
IHC-P
1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For the unpurified version use 1/1000- 1/10000 dilution |
靶标
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功能
In complex with TSC2, inhibits the nutrient-mediated or growth factor-stimulated phosphorylation of S6K1 and EIF4EBP1 by negatively regulating mTORC1 signaling. Seems not to be required for TSC2 GAP activity towards RHEB. Implicated as a tumor suppressor. Involved in microtubule-mediated protein transport, but this seems to be due to unregulated mTOR signaling. -
组织特异性
Highly expressed in skeletal muscle, followed by heart, brain, placenta, pancreas, lung, liver and kidney. Also expressed in embryonic kidney cells. -
疾病相关
Defects in TSC1 are the cause of tuberous sclerosis type 1 (TSC1) [MIM:191100]. It is an autosomal dominant multi-system disorder that affects especially the brain, kidneys, heart, and skin. TS1C is characterized by hamartomas (benign overgrowths predominantly of a cell or tissue type that occurs normally in the organ) and hamartias (developmental abnormalities of tissue combination). Clinical symptoms can range from benign hypopigmented macules of the skin to profound mental retardation with intractable seizures to premature death from a variety of disease-associated causes.
Defects in TSC1 may be a cause of focal cortical dysplasia of Taylor balloon cell type (FCDBC) [MIM:607341]. FCDBC is a subtype of cortical displasias linked to chronic intractable epilepsy. Cortical dysplasias display a broad spectrum of structural changes, which appear to result from changes in proliferation, migration, differentiation, and apoptosis of neuronal precursors and neurons during cortical development. -
结构域
The C-terminal putative coiled-coil domain is necessary for interaction with TSC2. -
翻译后修饰
Phosphorylation at Ser-505 does not affect interaction with TSC2. Phosphorylated upon DNA damage, probably by ATM or ATR. -
细胞定位
Cytoplasm. Membrane. At steady state found in association with membranes. - Information by UniProt
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数据库链接
- Entrez Gene: 7248 Human
- Omim: 605284 Human
- SwissProt: Q92574 Human
- Unigene: 370854 Human
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别名
- Hamartin antibody
- kiaa0243 antibody
- LAM antibody
see all
图片
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All lanes : Anti-Hamartin antibody [EP318Y] (ab40872) at 1/1000 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : TSC1 knockout MCF7 cell lysate
Lane 3 : Wild-type A549 cell lysate
Lane 4 : TSC1 knockout A549 cell lysate
Lane 5 : Wild-type HAP1 cell lysate
Lane 6 : TSC1 knockout HAP1 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 150 kDa
Observed band size: 130-150 kDa why is the actual band size different from the predicted?Western blot: Anti-TSC1 antibody [EP318Y] (ab40872) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab40872 was shown to bind specifically to TSC1. A band was observed at 130-150 kDa in wild-type MCF7 cell lysates with no signal observed at this size in TSC1 knockout cell line. To generate this image, wild-type and TSC1 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 (20 µg)
Lane 2: Hamartin knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Human skeletal muscle tissue lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab40872 observed at 150 kDa. Red - loading control, ab8245, observed at 37 kDa.ab40872 was shown to recognize Hamartin when Hamartin knockout samples were used, along with additional cross-reactive bands. Wild-type and Hamartin knockout samples were subjected to SDS-PAGE. ab40872 and ab8245 (loading control to GAPDH) were diluted 1/5000 and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.
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Anti-Hamartin antibody [EP318Y] (ab40872) at 1/1000 dilution (Purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG (H+L), Peroxidase conjugated)
Predicted band size: 150 kDaWe are not sure about the nature of the extra band.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hamartin antibody [EP318Y] (ab40872)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling Hamartin with purified ab40872 at 1/200 dilution (1.08 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Hamartin with Purified ab40872 at 1/20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488 ,ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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 (3)
ab40872 被引用在 3 文献中.
- Wang Y et al. miR-130a upregulates mTOR pathway by targeting TSC1 and is transactivated by NF-?B in high-grade serous ovarian carcinoma. Cell Death Differ 24:2089-2100 (2017). PubMed: 28800130
- Platt FM et al. Spectrum of phosphatidylinositol 3-kinase pathway gene alterations in bladder cancer. Clin Cancer Res 15:6008-17 (2009). PubMed: 19789314
- Pymar LS et al. Bladder tumour-derived somatic TSC1 missense mutations cause loss of function via distinct mechanisms. Hum Mol Genet 17:2006-17 (2008). PubMed: 18397877