Sample:
Muscle (Mouse) Lysate at 40 ug
Primary: Anti-Phospho-Smad1 (Ser463) (bs-10381R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 51 kD
Observed band size: 51 kD
- ￥1180 - 2800
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- bs-10381R
- 2025年10月24日
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50ul/100ul/200ul
| 规格： | 50ul | 产品价格： | ￥1180.0 |
|---|---|---|---|
| 规格： | 100ul | 产品价格： | ￥1980.0 |
| 规格： | 200ul | 产品价格： | ￥2800.0 |
| 产品编号 | bs-10381R |
| 英文名称 | phospho-Smad1 (Ser463) Rabbit pAb |
| 中文名称 | 磷酸化细胞信号转导分子Smad-1抗体 |
| 英文别名 | Smad1(phospho S463); Smad1(phospho Ser463); p-Smad1(Ser463); Mothers against decapentaplegic homolog 1; BSP 1; BSP1; BSP-1; Dwarfin A; DwfA; hSMAD 1; hSMAD1; JV 41; JV4 1; JV4-1; JV41; MAD mothers against decapentaplegic homolog 1; Mad related protein 1; MADH 1; MADH1; Madr 1; MADR1; Mothers against decapentaplegic homolog 1; Mothers against DPP homolog 1; Sma and Mad related protein 1; SMAD 1; SMAD-1; SMAD family member 1; SMAD mothers against DPP homolog 1 Drosophila; Spinal muscular atrophy distal with upper limb predominance; TGF beta signaling protein 1; Transforming growth factor beta signaling protein 1; SMAD1_HUMAN; MAD homolog 1; Mothers against DPP homolog 1; Mad-related protein 1; SMAD family member 1; Smad1; Transforming growth factor-beta-signaling protein 1. |
| 产品应用 | WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=2ug/Test Not yet tested in other applications. |
| 交叉反应 | Human, Mouse, Rat, Hela (Pig) |
| 抗体来源 | Rabbit |
| 免疫原 | KLH conjugated Synthesised phosphopeptide derived from human Smad1 around the phosphorylation site of Ser463 |
| 亚型 | IgG |
| 性状 | Liquid |
| 纯化方法 | affinity purified by Protein A |
| 克隆类型 | Polyclonal |
| 理论分子量 | 51 kDa |
| 浓度 | 1mg/ml |
| 储存液 | 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. |
| 研究领域 | Cancer > Growth factors > TGF Epigenetics and Nuclear Signaling > Nuclear Signaling Pathways > SMADs Signal Transduction > Signaling Pathway > Nuclear Signaling > SMADs Stem Cells > Signaling Pathways > TGF beta > Cytoplasmic |
| 亚基 | Interacts with HGS, NANOG and ZCCHC12 (By similarity). May form trimers with another SMAD1 and the co-SMAD SMAD4. Interacts with PEBP2-alpha subunit, CREB-binding protein (CBP), p300, SMURF1, SMURF2 and HOXC8. Associates with ZNF423 or ZNF521 in response to BMP2 leading to activate transcription of BMP target genes. Interacts with SKOR1. Interacts (via MH2 domain) with LEMD3. Binding to LEMD3 results in at least a partial reduction of receptor-mediated phosphorylation. Forms a ternary complex with PSMB4 and OAZ1 before PSMB4 is incorporated into the 20S proteasome. |
| 亚细胞定位 | Cytoplasm. Nucleus. Note=Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4. Co-localizes with LEMD3 at the nucleus inner membrane. |
| 组织特异性 | Ubiquitous. Highest expression seen in the heart and skeletal muscle. |
| 翻译后修饰 | Phosphorylated on serine by BMP type 1 receptor kinase.
Ubiquitin-mediated proteolysis by SMAD-specific E3 ubiquitin ligase SMURF1. |
| 相似性 | Belongs to the dwarfin/SMAD family. Contains 1 MH1 (MAD homology 1) domain. Contains 1 MH2 (MAD homology 2) domain. |
| 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. |
| 保存条件 | Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. |
| 注意事项 | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. |
| 背景资料 | The protein encoded by this gene belongs to the SMAD, a family of proteins similar to the gene products of the Drosophila gene 'mothers against decapentaplegic' (Mad) and the C. elegans gene Sma. SMAD proteins are signal transducers and transcriptional modulators that mediate multiple signaling pathways. This protein mediates the signals of the bone morphogenetic proteins (BMPs), which are involved in a range of biological activities including cell growth, apoptosis, morphogenesis, development and immune responses. In response to BMP ligands, this protein can be phosphorylated and activated by the BMP receptor kinase. The phosphorylated form of this protein forms a complex with SMAD4, which is important for its function in the transcription regulation. This protein is a target for SMAD-specific E3 ubiquitin ligases, such as SMURF1 and SMURF2, and undergoes ubiquitination and proteasome-mediated degradation. Alternatively spliced transcript variants encoding the same protein have been observed. [provided by RefSeq]. |
| 应用 | 推荐稀释比例 |
| {WB} | {1:500-2000} |
| {IHC-P} | {1:100-500} |
| {IHC-F} | {1:100-500} |
| {IF} | {1:100-500} |
| {Flow-Cyt} | {2ug/Test} |
Paraformaldehyde-fixed, paraffin embedded (human brain glioma); Antigen retrieval by microwave in sodium citratebuffer (pH6.0) ; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (3%BSA) at RTfor 30min; Antibody incubation with (Smad1 (Ser463)) Polyclonal/MonoclonalAntibody, Unconjugated (bs-10381R) at 1:400 overnight at 4℃,followed by conjugation to the secondary antibody (labeled with HRP)and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat heart); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-Smad1 (Ser463)) Polyclonal Antibody, Unconjugated (bs-10381R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.
Sample:
Heart (Mouse) Lysate at 40 ug
Primary: Anti- Phospho-Smad1 (Ser463) (bs-10381R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 51 kD
Observed band size: 51 kD
Heart (Mouse) Lysate at 40 ug
Primary: Anti- Phospho-Smad1 (Ser463) (bs-10381R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 51 kD
Observed band size: 51 kD
Blank control（black line）:Hela.
Primary Antibody (green line): Rabbit Anti-Phospho-Smad1 (Ser463) antibody (bs-10381R)
Dilution:2ug/Test;
Secondary Antibody（white blue line）: Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
Isotype control（orange line）: Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Primary Antibody (green line): Rabbit Anti-Phospho-Smad1 (Ser463) antibody (bs-10381R)
Dilution:2ug/Test;
Secondary Antibody（white blue line）: Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
Isotype control（orange line）: Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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at 10, 25, and 50 μM). At 24 h post-treatment, celllysates were obtained; SDS-PAGE and Western blotting were performed as described in Section 3, Methods. AR-A014418dose-dependent decreases of phospho-β-catenin (Ser33/37) and phospho-glycogen synthase
(Ser473)(193H12)Rabbit mAb，1:300,稀释，二抗是驴抗兔，1：2000，我是实验室的新手，刚呆俩月，受 此挫折，一头雾水，请过来人或者有经验的同学帮忙解答怎样解决，非常感谢！ 碧峤 pAkt有两个位点，Ser473和Thr308，其中Ser473相对容易显色。如果显色失败，可能的原因:1 上样量过少，我们一般都是30μg；2 一抗孵育时间过短，我们一般24h~48h；3 一抗是否工作有待排除 米宝
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phospho-Smad1 (Ser463) Rabbit pAb(bs-10381R)-50ul/100ul/200ul
￥1180 - 2800
