重组Anti-STK3/MST-2抗体[EP1466Y] (ab52641)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1466Y] to STK3/MST-2
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-STK3/MST-2抗体[EP1466Y]
参阅全部 STK3/MST-2 一抗 -
描述
兔单克隆抗体[EP1466Y] to STK3/MST-2 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human STK3/MST-2 aa 1-100 (N terminal). The exact sequence is proprietary.
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阳性对照
- ICC/IF: Wildtype HAP1 cells, NIH/3T3 (Mouse embryonic fibroblast) cells; WB: NIH/3T3, Hek293, HeLa, C6 cell lysate; IHC-P: Human lymphoma tissue; Flow Cyt (intra): NIH/3T3 (Mouse embryonic fibroblast).
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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. Stable for 12 months at -20°C. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP1466Y -
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
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab52641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) |
1/70.
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ICC/IF |
Use a concentration of 5 µg/ml.
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WB |
1/10000 - 1/50000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa).
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IP |
1/50 - 1/100.
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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.
Antigen retrieval step strongly recommended for enhanced signal. |
说明 |
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Flow Cyt (Intra)
1/70. |
ICC/IF
Use a concentration of 5 µg/ml. |
WB
1/10000 - 1/50000. Detects a band of approximately 56 kDa (predicted molecular weight: 56 kDa). |
IP
1/50 - 1/100. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Antigen retrieval step strongly recommended for enhanced signal. |
靶标
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功能
Stress-activated, pro-apoptotic kinase which, following caspase-cleavage, enters the nucleus and induces chromatin condensation followed by internucleosomal DNA fragmentation. Key component of the Hippo signaling pathway which plays a pivotal role in organ size control and tumor suppression by restricting proliferation and promoting apoptosis. The core of this pathway is composed of a kinase cascade wherein MST1/MST2, in complex with its regulatory protein SAV1, phosphorylates and activates LATS1/2 in complex with its regulatory protein MOB1, which in turn phosphorylates and inactivates YAP1 oncoprotein and WWTR1/TAZ. Phosphorylation of YAP1 by LATS2 inhibits its translocation into the nucleus to regulate cellular genes important for cell proliferation, cell death, and cell migration. MST1/MST2 are required to repress proliferation of mature hepatocytes, to prevent activation of facultative adult liver stem cells (oval cells), and to inhibit tumor formation. Phosphorylates NKX2-1. -
组织特异性
Expressed at high levels in adult kidney, skeletal and placenta tissues and at very low levels in adult heart, lung and brain tissues. -
序列相似性
Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. STE20 subfamily.
Contains 1 protein kinase domain.
Contains 1 SARAH domain. -
细胞定位
Cytoplasm. Nucleus. The caspase-cleaved form cycles between nucleus and cytoplasm. - Information by UniProt
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数据库链接
- Entrez Gene: 6788 Human
- Entrez Gene: 56274 Mouse
- Entrez Gene: 65189 Rat
- Omim: 605030 Human
- SwissProt: Q13188 Human
- SwissProt: Q9JI10 Mouse
- SwissProt: O54748 Rat
- Unigene: 492333 Human
see all -
别名
- 0610042I06Rik antibody
- EC 2.7.11.1 antibody
- FLJ90748 antibody
see all
图片
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: STK3/MST-2 knockout HAP1 cell lysate (20 µg)
Lane 3: NIH/3T3 cell lysate (20 µg)
Lane 4: 293T cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab52641 observed at 56 kDa. Red - loading control, ab8245, observed at 37 kDa.ab52641 was shown to specifically react with STK3/MST-2 when STK3/MST-2 knockout samples were used. Wild-type and STK3/MST-2 knockout samples were subjected to SDS-PAGE. ab52641 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 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 1/10 000 dilution for 1 h at room temperature before imaging.
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ab52641 staining STK3/MST-2 in wild-type HAP1 cells (top panel) and STK3/MST-2 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab52641 at 5μg/ml concentration and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
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Immunohistochemical staining of paraffin embedded human breast carcinoma with purified ab52641 at a working dilution of 1/50. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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NIH/3T3 (Mouse embryonic fibroblast) cells were fixed with4% paraformaldehyde and permeabilised with 90% methanol. The primary antibody (ab52641) was used at a 1/70 dilution (1 µg) (red). A Goat anti rabbit IgG (Alexa Fluorr® 488, ab150077) was used as the secondary antibody at a 1/2000 dilution. A Rabbit monoclonal IgG (ab172730) (black) was used as an isotype control.Cells without incubation with primary antibody and secondary antibody were used as an unlabelled control (blue).
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ab52641 (purified) at 1/50 immunoprecipitating STK3/MST-2 in 10 μg C6 cell lysate (Lanes 1 and 2, observed at 56 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, HRP Veriblot for IP (ab131366) was used for detection (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
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Confocal image showing cytoplasmic staining of STK3/MST-2 in NIH/3T3 (mouse embryonic fibroblast) cells using ab52641 . The cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. The cells were then incubated with ab52641 at 1/70 dilution and counterstained with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/100 dilution (red). Goat anti Rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at 1/1000 dilution (green). Nuclei counterstained with DAPI (blue).
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All lanes : Anti-STK3/MST-2 antibody [EP1466Y] (ab52641) at 1/50000 dilution (purified)
Lane 1 : C6 whole cell lysate
Lane 2 : NIH/3T3 whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
All lanes : Anti-STK3/MST-2 antibody [EP1466Y] (ab52641) at 1/50000 dilution (purified)
Lane 1 : HEK293 whole cell lysate
Lane 2 : HeLa whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 56 kDa
Observed band size: 56 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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 (33)
ab52641 被引用在 33 文献中.
- Ahmadzai MM et al. LPAR1 regulates enteric nervous system function through glial signaling and contributes to chronic intestinal pseudo-obstruction. J Clin Invest 132:N/A (2022). PubMed: 35166239
- Tsaridou S et al. 53BP1-mediated recruitment of RASSF1A to ribosomal DNA breaks promotes local ATM signaling. EMBO Rep 23:e54483 (2022). PubMed: 35758159
- García-Gutiérrez L et al. Interaction of LATS1 with SMAC links the MST2/Hippo pathway with apoptosis in an IAP-dependent manner. Cell Death Dis 13:692 (2022). PubMed: 35941108
- Romano D et al. Proteasomal down-regulation of the proapoptotic MST2 pathway contributes to BRAF inhibitor resistance in melanoma. Life Sci Alliance 5:N/A (2022). PubMed: 36038253
- Drexler R et al. The clinical relevance of the Hippo pathway in pancreatic ductal adenocarcinoma. J Cancer Res Clin Oncol 147:373-391 (2021). PubMed: 33098447