重组Anti-Smad1 (phospho S463 + S465)抗体[EPR20662-20] (ab226821)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20662-20] to Smad1 (phospho S463 + S465)
- Suitable for: WB, Dot blot, ICC/IF, IP
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-Smad1 (phospho S463 + S465)抗体[EPR20662-20]
参阅全部 Smad1 一抗 -
描述
兔单克隆抗体[EPR20662-20] to Smad1 (phospho S463 + S465) -
宿主
Rabbit -
特异性
Based on sequence homology this antibody also reacts with Smad5 (phospho S463/S465) and Smad9 (phospho S465/S467). -
经测试应用
适用于: WB, Dot blot, ICC/IF, IPmore details -
种属反应性
与反应: Mouse, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa grown in serum-free media overnight, treated with 100 ng/ml Calyculin A (ab141784) for 15min followed by Calyculin A removal and treatment with 100 ng/ml BMP2 for 30min, whole cell lysate; NIH/3T3 cultured in serum-free media overnight then treated with 50 ng/ml BMP2 for 30min whole cell lysate. ICC/IF: NIH3T3 cells FBS-deprived overnight before treatment with 50 ng/ml hBMP2 for 30min. IP: NIH/3T3 grown in serum-free media overnight then treated with 50 ng/ml BMP2 for 30min whole cell lysate.
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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, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR20662-20 -
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
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab226821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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WB |
1/1000. Detects a band of approximately 60 kDa (predicted molecular weight: 52 kDa).
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Dot blot |
1/1000.
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ICC/IF |
1/100.
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IP |
1/30.
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说明 |
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WB
1/1000. Detects a band of approximately 60 kDa (predicted molecular weight: 52 kDa). |
Dot blot
1/1000. |
ICC/IF
1/100. |
IP
1/30. |
靶标
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功能
Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD1 is a receptor-regulated SMAD (R-SMAD). SMAD1/OAZ1/PSMB4 complex mediates the degradation of the CREBBP/EP300 repressor SNIP1. -
组织特异性
Ubiquitous. Highest expression seen in the heart and skeletal muscle. -
序列相似性
Belongs to the dwarfin/SMAD family.
Contains 1 MH1 (MAD homology 1) domain.
Contains 1 MH2 (MAD homology 2) domain. -
翻译后修饰
Phosphorylated on serine by BMP type 1 receptor kinase.
Ubiquitin-mediated proteolysis by SMAD-specific E3 ubiquitin ligase SMURF1. -
细胞定位
Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4. Co-localizes with LEMD3 at the nucleus inner membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 4086 Human
- Entrez Gene: 17125 Mouse
- Omim: 601595 Human
- SwissProt: Q15797 Human
- SwissProt: P70340 Mouse
- Unigene: 604588 Human
- Unigene: 223717 Mouse
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别名
- BSP-1 antibody
- BSP1 antibody
- HsMAD1 antibody
see all
图片
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All lanes : Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (ab226821) at 1/1000 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) grown in serum-free media overnight, whole cell lysate
Lane 2 : HeLa grown in serum-free media overnight, then treated with 100 ng/ml Calyculin A (ab141784) for 15 minutes, followed by Calyculin A removal and treatment with 100 ng/ml BMP2 for 30 minutes, whole cell lysate
Lane 3 : NIH/3T3 (mouse embryo fibroblast cell line) grown in serum-free media overnight, whole cell lysate
Lane 4 : NIH/3T3 cultured in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 52 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Exposure time:
Lanes 1 and 2: 3 minutes.
Lanes 3 and 4: 30 seconds.Blocking/Dilution buffer: 5% NFDM/TBST.
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Dot blot analysis of Smad1 (phospho S463 + S465) labeled with ab226821 at 1/1000 dilution.
Lane 1: Smad1 (phospho S463/S465) peptide;
Lane 2: Smad1 (phospho S463) peptide;
Lane 3: Smad1 (phospho S465) peptide;
Lane 4: Smad1 peptide (not phosphorylated);
Lane 5: Smad5 (phospho S463/S465) peptide;
Lane 6: Smad5 (phospho S463) peptide;
Lane 7: Smad5 (phospho S465) peptide;
Lane 9: Smad5 peptide (not phosphorylated).
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
Based on sequence homology, this antibody cross-reacts with Smad5 (phospho S463/S465) and Smad9 (phospho S465/S467).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling Smad1 (phospho S463 + S465) with ab226821 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Nuclear staining in hBMP2-treated NIH/3T3 cells. Cells were FBS-deprived overnight before treatment with 50 ng/ml hBMP2 for 30 minutes.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Smad 1 (phospho S463 + S465) was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryo fibroblast cell line) grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate with ab226821 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab226821 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: NIH/3T3 grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate 10 μg (Input).
Lane 2: ab226821 IP in NIH/3T3 grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab226821 in NIH/3T3 grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
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
Certificate of Compliance
文献 (4)
ab226821 被引用在 4 文献中.
- Tang C et al. Omentin‑1 induces osteoblast viability and differentiation via the TGF‑β/Smad signaling pathway in osteoporosis. Mol Med Rep 25:N/A (2022). PubMed: 35179221
- Zhang Y et al. CellCall: integrating paired ligand-receptor and transcription factor activities for cell-cell communication. Nucleic Acids Res 49:8520-8534 (2021). PubMed: 34331449
- Zhang N et al. Inhibiting microRNA-424 in bone marrow mesenchymal stem cells-derived exosomes suppresses tumor growth in colorectal cancer by upregulating TGFBR3. Arch Biochem Biophys 709:108965 (2021). PubMed: 34129838
- Chen L et al. H2 O2 induces oxidative stress damage through the BMP-6/SMAD/hepcidin axis. Dev Growth Differ 62:139-146 (2020). PubMed: 32012242