重组Anti-active YAP1抗体[EPR19812] (ab205270)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19812] to active YAP1
- Suitable for: ICC/IF, IHC-P, WB
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-active YAP1抗体[EPR19812]
参阅全部 active YAP1 一抗 -
描述
兔单克隆抗体[EPR19812] to active YAP1 -
宿主
Rabbit -
特异性
ab205270 is specific to the active (non-phosphorylated) form of YAP1. -
经测试应用
适用于: ICC/IF, IHC-P, WBmore details -
种属反应性
与反应: Mouse, Human -
免疫原
This product was produced with the following immunogens:
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers. -
阳性对照
- WB: 293A cell lysate serum starved overnight, then 10% FBS was added to medium for 1 hour; 293A cell lysate serum starved overnight and then treated with Lambda phosphatase lysate. Human kidney and skin lysates and mouse testis and skin lysates. HaCaT whole cell lysate, 293A cell lysate. HeLa and NIH/3T3 treated with Calyculin A. IHC-P: Human breast and breast cancer tissues; Mouse skin tissue. ICC/IF: 293A cell line serum starved overnight, then 10% FBS was added to medium for 1 hour; 293A 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: 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
EPR19812 -
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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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab205270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 |
---|---|---|
ICC/IF | (2) |
1/500.
|
IHC-P | (11) |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
WB | (2) |
1/1000. Detects a band of approximately 75 kDa (predicted molecular weight: 54 kDa).
|
说明 |
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ICC/IF
1/500. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 75 kDa (predicted molecular weight: 54 kDa). |
靶标
- Information by UniProt
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数据库链接
- Entrez Gene: 10413 Human
- Entrez Gene: 22601 Mouse
- SwissProt: P46937 Human
- SwissProt: P46938 Mouse
- Unigene: 503692 Human
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别名
- Protein yorkie homolog antibody
- Transcriptional coactivator YAP1 antibody
- YAP1 antibody
see all
图片
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All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1 µg/ml
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : YAP1 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 54 kDaLanes 1 - 3: Merged signal (red and green). Green - ab205270 observed at 54 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab205270 was shown to recognize active YAP1 in wild-type HAP1 cells as signal was lost at the expected MW in active YAP1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and active YAP1 knockout samples were subjected to SDS-PAGE. Ab205270 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemical analysis of paraffin-embedded human breast tissue labeling active YAP1 with ab205270 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Mainly nuclear staining on human breast is observed [PMID: 18617895].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Ab205270 staining active YAP1 in HUVEC (human umbilical vein endothelial cell) cells by Immunocytochemistry/Immunofluorescence (ICC/IF). The cells were fixed 4% Paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody at 1:500 dilution. An Alexa Fluor® 488 Goat anti-Rabbit was used as a secondary antibody at 1:1000 dilution. DAPI was used as a nuclear counter stain. Confocal image showing nuclear and cytoplasmic staining in HUVEC cells.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton/PBS permeabilized (RT, 5 mins) 293A (Human epithelial cell line from embryonic kidney transformed with sheared human adenovirus type 5 DNA) cells labeling active YAP1 with ab205270 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 594) secondary antibody at 1/1000 dilution.
The images showed weak staining on 293A cells under serum starvation overnight. After 10% FBS was added to the medium for 1h, the nuclear staining was increased.
The data was kindly provided by our collaborator Dr. Bin Zhao (Zhejiang University).
The nuclear counterstain is DAPI (blue). Counterstained with Phalloidin-technology® Alexa Fluor 488 at 1/1000 dilution.
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All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1/1000 dilution
Lane 1 : Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg
Lane 2 : HeLa treated with 100ng/ml Calyculin A for 30 min whole cell lysate at 15 µg
Lane 3 : HeLa treated with 100ng/ml Calyculin A for 30 min whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 54 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 80 secondsBlocking buffer: 5% NFDM/TBST.
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All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1/1000 dilution
Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 15 µg
Lane 2 : NIH/3T3 treated with 100ng/ml Calyculin A for 30 min whole cell lysate at 15 µg
Lane 3 : NIH/3T3 treated with 100ng/ml Calyculin A for 30 min whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 54 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 20 secondsBlocking buffer: 5% NFDM/TBST.
-
All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1/1000 dilution
Lane 1 : 293A cells under serum starvation overnight lysate
Lane 2 : 293A cell lysate under serum starvation overnight, then 10% FBS was added to medium for 1 hour
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 54 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
Serum starvation induces active YAP1 Ser127 phosphorylation. The level of active YAP1 protein is inversely proportional to p-YAP1 Ser127 level (PMID: 22884261).
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All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1/1000 dilution
Lane 1 : Human kidney lysate
Lane 2 : Human skin lysate
Lane 3 : HaCaT (Human keratinocyte cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
Lanes 1-2 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Lane 3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 54 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/2: 8 seconds; Lane 3: 30 seconds.
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All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1/1000 dilution
Lane 1 : Mouse testis lysate
Lane 2 : Mouse skin lysate
Lane 3 : Mouse liver lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 54 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 10 seconds; Lane 2: 8 seconds; Lane 3: 3 minutes.
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All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1/1000 dilution
Lane 1 : 293A cell lysate serum starved overnight and then 10% FBS added to the medium for 1 hour
Lane 2 : 293A cell lysate serum starved overnight
Lane 3 : 293A cell lysate serum starved overnight and then treated with Lambda phosphatase
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : donkey anti-rabbit at 1/1000 dilution
Predicted band size: 54 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% BSA/TBST.
The two panels for pYAPS127 were just shorter and longer exposure.
The data was kindly provided by our collaborator Dr. Bin Zhao (Zhejiang University).
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All lanes : Anti-active YAP1 antibody [EPR19812] (ab205270) at 1/1000 dilution
Lane 1 : 293A cell lysate
Lane 2 : YAP/TAZ knockout 293A cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : donkey anti-rabbit at 1/1000 dilution
Predicted band size: 54 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% BSA/TBST.
The data was kindly provided by our collaborator Dr. Bin Zhao (Zhejiang University).
-
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling active YAP1 with ab205270 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and cytoplasmic staining on human breast cancer is observed [PMID: 24559095].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunohistochemical analysis of paraffin-embedded mouse skin tissue labeling active YAP1 with ab205270 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Mainly nuclear staining on mouse skin is observed [PMID: 21610251].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton/PBS permeabilized (RT, 5 mins) 293A (Human epithelial cell line from embryonic kidney transformed with sheared human adenovirus type 5 DNA) cells labeling active YAP1 with ab205270 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 594) secondary antibody at 1/1000 dilution.
The images showed nuclear staining on 293A cells, and background staining on YAP/TAZ knockout 293A cells.
The data was kindly provided by our collaborator Dr. Bin Zhao (Zhejiang University).
The nuclear counterstain is DAPI (blue). Counterstained with Phalloidin-technology® Alexa Fluor 488 at 1/1000 dilution.
-
Immunohistochemical analysis of paraffin-embedded human liver tissue labeling active YAP1 with ab205270 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Negative control: no staining on human liver [PMID:17974916].
Counter stained with Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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 (70)
ab205270 被引用在 70 文献中.
- Dai J et al. Astrocytic laminin-211 drives disseminated breast tumor cell dormancy in brain. Nat Cancer 3:25-42 (2022). PubMed: 35121993
- Wu Q et al. YAP signaling in horizontal basal cells promotes the regeneration of olfactory epithelium after injury. Stem Cell Reports 17:664-677 (2022). PubMed: 35148842
- Ke WL et al. m6A demethylase FTO regulates the apoptosis and inflammation of cardiomyocytes via YAP1 in ischemia-reperfusion injury. Bioengineered 13:5443-5452 (2022). PubMed: 35176940
- He J et al. A Novel Small Molecular Prostaglandin Receptor EP4 Antagonist, L001, Suppresses Pancreatic Cancer Metastasis. Molecules 27:N/A (2022). PubMed: 35208999
- Zang J et al. MiR-326 inhibits trophoblast growth, migration, and invasion by targeting PAX8 via Hippo pathway. Reprod Biol Endocrinol 20:38 (2022). PubMed: 35209928