重组Anti-Tuberin抗体[Y320] (ab32554)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y320] to Tuberin
- Suitable for: Flow Cyt (Intra), WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Tuberin抗体[Y320]
参阅全部 Tuberin 一抗 -
描述
兔单克隆抗体[Y320] to Tuberin -
宿主
Rabbit -
特异性
This antibody recognises Tuberous sclerosis complex-2 (TCS2 also known as Tuberin. It is predicted to detect isoform 2-4 based on sequence homology.
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经测试应用
适用于: Flow Cyt (Intra), WB, IHC-Pmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human Tuberin aa 1750-1850 (C terminal). The exact sequence is proprietary.
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阳性对照
- WB: HAP1 and HeLa whole cell lysates. Rat and mouse brain tissue lysates; Flow Cyt (intra): Jurkat cells; IHC: Rat and mouse liver tissue. Human lung carcinoma tissue.
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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. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
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
Y320 -
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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Positive Controls
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32554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) |
1/30.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 dilution |
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WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 220 kDa (predicted molecular weight: 200 kDa).
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IHC-P |
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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说明 |
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Flow Cyt (Intra)
1/30. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 dilution |
WB
1/1000 - 1/10000. Detects a band of approximately 220 kDa (predicted molecular weight: 200 kDa). |
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
靶标
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功能
In complex with TSC1, inhibits the nutrient-mediated or growth factor-stimulated phosphorylation of S6K1 and EIF4EBP1 by negatively regulating mTORC1 signaling. Acts as a GTPase-activating protein (GAP) for the small GTPase RHEB, a direct activator of the protein kinase activity of mTORC1. Implicated as a tumor suppressor. Involved in microtubule-mediated protein transport, but this seems to be due to unregulated mTOR signaling. Stimulates weakly the intrinsic GTPase activity of the Ras-related proteins RAP1A and RAB5 in vitro. Mutations in TSC2 lead to constitutive activation of RAP1A in tumors. -
组织特异性
Liver, brain, heart, lymphocytes, fibroblasts, biliary epithelium, pancreas, skeletal muscle, kidney, lung and placenta. -
疾病相关
Defects in TSC2 are the cause of tuberous sclerosis type 2 (TSC2) [MIM:613254]. TSC2 is an autosomal dominant multi-system disorder that affects especially the brain, kidneys, heart, and skin. It 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 TSC2 are a cause of lymphangioleiomyomatosis (LAM) [MIM:606690]. LAM is a progressive and often fatal lung disease characterized by a diffuse proliferation of abnormal smooth muscle cells in the lungs. It affects almost exclusively young women and can occur as an isolated disorder or in association with tuberous sclerosis complex. -
序列相似性
Contains 1 Rap-GAP domain. -
翻译后修饰
Phosphorylation at Ser-1387, Ser-1418 or Ser-1420 does not affect interaction with TSC1.
Phosphorylation at Ser-939 and Thr-1462 by PKB/AKT1 is induced by growth factor stimulation. -
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: 7249 Human
- Entrez Gene: 22084 Mouse
- Entrez Gene: 24855 Rat
- Omim: 191092 Human
- SwissProt: P49815 Human
- SwissProt: Q61037 Mouse
- SwissProt: P49816 Rat
- Unigene: 90303 Human
see all -
别名
- FLJ43106 antibody
- LAM antibody
- OTTHUMP00000158940 antibody
see all
图片
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All lanes : Anti-Tuberin antibody [Y320] (ab32554) at 1/5000 dilution (purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate boiled
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate unboiled
Lane 3 : Mouse brain lysate boiled
Lane 4 : Mouse brain lysate unboiled
Lane 5 : Rat brain lysate boiled
Lane 6 : Rat brain lysate unboiled
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 200 kDa
Observed band size: 200 kDaThe unboiled lysate are recommended to be used in SDS-PAGE, in order to prevent membrane protein aggregation.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling Tuberin with purified ab32554 at 1/100 dilution (2.97 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling Tuberin with purified ab32554 at 1/100 dilution (2.97 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling Tuberin with purified ab32554 at 1/100 dilution (2.97 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). 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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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Tuberin knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - unpurified ab32554 observed at 180 kDa. Red - loading control, ab18058, observed at 130 kDa.ab32554 was shown to specifically react with tuberin in wild-type HAP1 cells. No band was observed when tuberin knockout samples were used. Wild-type and tuberin knockout samples were subjected to SDS-PAGE. Ab32554 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed 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 1/10000 dilution for 1 hour at room temperature before imaging.
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Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling Tuberin with purified ab32554 at 1/30 dilution (10µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr® 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 (18)
ab32554 被引用在 18 文献中.
- Shiomi E et al. Relationship between miR-155 expression and clear cell papillary renal cell carcinoma in the dialyzed kidney. IJU Case Rep 4:127-131 (2021). PubMed: 33977237
- Zhang X et al. Tanshinone IIA protects against heart failure post-myocardial infarction via AMPKs/mTOR-dependent autophagy pathway. Biomed Pharmacother 112:108599 (2019). PubMed: 30798134
- Wang H et al. Feedback Activation of SGK3 and AKT Contributes to Rapamycin Resistance by Reactivating mTORC1/4EBP1 Axis via TSC2 in Breast Cancer. Int J Biol Sci 15:929-941 (2019). PubMed: 31182914
- Hadley G et al. Investigation of the novel mTOR inhibitor AZD2014 in neuronal ischemia. Neurosci Lett 706:223-230 (2019). PubMed: 31100427
- Ruvolo PP et al. Role of MSC-derived galectin 3 in the AML microenvironment. Biochim Biophys Acta 1865:959-969 (2018). PubMed: 29655803