Tissue/cell: rat kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Smad7/Smad6 Polyclonal Antibody, Unconjugated(bs-0071R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
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- 规格：
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-0566R |
| 英文名称 | SMAD7 Rabbit pAb |
| 中文名称 | Smad7抗体 |
| 英文别名 | MAD(mothers against decapentaplegic Drosophila) homolog 7; MAD; Mad homolog 7; MAD mothers against decapentaplegic homolog 7; MADH 7; MADH 8; MADH8; Mothers Against Decapentaplegic Drosophila Homolog of 7; Mothers against decapentaplegic homolog 7; Mothers against decapentaplegic homolog 8; Mothers against DPP homolog 7; Mothers against DPP homolog 8; SMA-AND MAD-RELATED PROTEIN 7; SMAD 7; SMAD; SMAD family member 7; SMAD, mothers against DPP homolog 7(Drosophila); SMAD, mothers against DPP homolog 7; SMAD7_HUMAN. |
| 产品应用 | WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100-500, IF=1:100-500, Flow-Cyt=1ug/Test Not yet tested in other applications. |
| 交叉反应 | Human, Mouse, Rat (Pig, Cow) |
| 抗体来源 | Rabbit |
| 免疫原 | KLH conjugated synthetic peptide derived from human Smad7 |
| 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 | 46 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 | Epigenetics and Nuclear Signaling > Nuclear Signaling Pathways > SMADs Signal Transduction > Signaling Pathway > Nuclear Signaling > SMADs |
| 亚基 | Interacts with WWP1. Interacts with COPS5. Interacts with NEDD4L. Interacts with STAMBP. Interacts with RNF111, AXIN1 and AXIN2. Interacts with PPP1R15A. Interacts (via MH2 domain) with EP300. Interacts with ACVR1B, SMURF1, SMURF2 and TGFBR1; SMAD7 recruits SMURF1 and SMURF2 to the TGF-beta receptor and regulates its degradation. Interacts with PDPK1 (via PH domain). |
| 亚细胞定位 | Nucleus. Cytoplasm. Note=Interaction with NEDD4L or RNF111 or induces translocation from the nucleus to the cytoplasm. TGF-beta stimulates its translocation from the nucleus to the cytoplasm. PDPK1 inhibits its translocation from the nucleus to the cytoplasm in response to TGF-beta. |
| 组织特异性 | Ubiquitous with higher expression in the lung and vascular endothelium. |
| 翻译后修饰 | Phosphorylation on Ser-249 does not affect its stability, nuclear localization or inhibitory function in TGFB signaling; however it affects its ability to regulate transcription. Phosphorylated by PDPK1.
Ubiquitinated by WWP1 (By similarity). Polyubiquitinated by RNF111, which is enhanced by AXIN1 and promotes proteasomal degradation. In response to TGF-beta, ubiquitinated by SMURF1; which promotes its degradation. Acetylation prevents ubiquitination and degradation mediated by 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 | Antagonist of signaling by TGF-beta (transforming growth factor) type 1 receptor superfamily members; has been shown to inhibit TGF-beta (Transforming growth factor) and activin signaling by associating with their receptors thus preventing SMAD2 access. Functions as an adapter to recruit SMURF2 to the TGF-beta receptor complex. Also acts by recruiting the PPP1R15A-PP1 complex to TGFBR1, which promotes its dephosphorylation. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator. |
| 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 is a nuclear protein that binds the E3 ubiquitin ligase SMURF2. Upon binding, this complex translocates to the cytoplasm, where it interacts with TGF-beta receptor type-1 (TGFBR1), leading to the degradation of both the encoded protein and TGFBR1. Expression of this gene is induced by TGFBR1. Variations in this gene are a cause of susceptibility to colorectal cancer type 3 (CRCS3). Several transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jun 2010] |
| 应用 | 推荐稀释比例 |
| {WB} | {1:500-2000} |
| {IHC-P} | {1:100-500} |
| {IHC-F} | {1:100-500} |
| {ICC/IF} | {1:100-500} |
| {IF} | {1:100-500} |
| {Flow-Cyt} | {1ug/Test} |
Sample:
Lung(Mouse) Lysate at 30 ug
Placenta(Mouse) Lysate at 30 ug
Large intestine(Mouse) Lysate at 30 ug
Primary: Anti- MADH7/Smad7 (bs-0566R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 46 kD
Observed band size: 50 kD
Lung(Mouse) Lysate at 30 ug
Placenta(Mouse) Lysate at 30 ug
Large intestine(Mouse) Lysate at 30 ug
Primary: Anti- MADH7/Smad7 (bs-0566R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 46 kD
Observed band size: 50 kD
Paraformaldehyde-fixed, paraffin embedded (rat lung); 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 (Smad7) Polyclonal Antibody, Unconjugated (bs-0566R) at 1:600 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Blank control: SH-SY5Y.
Primary Antibody (green line): Rabbit Anti-MADH7/Smad7 antibody (bs-0566R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
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-MADH7/Smad7 antibody (bs-0566R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
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.
Sample:
Lane 1: Cerebrum (Mouse) Lysate at 40 ug
Lane 2: Cerebrum (Rat) Lysate at 40 ug
Lane 3: Stomach (Mouse) Lysate at 40 ug
Lane 4: Lung (Mouse) Lysate at 40 ug
Lane 5: Kidney (Mouse) Lysate at 40 ug
Primary: Anti-MADH7/Smad7 (bs-0566R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 50 kD
Observed band size: 50 kD
Lane 1: Cerebrum (Mouse) Lysate at 40 ug
Lane 2: Cerebrum (Rat) Lysate at 40 ug
Lane 3: Stomach (Mouse) Lysate at 40 ug
Lane 4: Lung (Mouse) Lysate at 40 ug
Lane 5: Kidney (Mouse) Lysate at 40 ug
Primary: Anti-MADH7/Smad7 (bs-0566R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 50 kD
Observed band size: 50 kD
Paraformaldehyde-fixed, paraffin embedded (rat stomach); 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 (MADH7) Polyclonal Antibody, Unconjugated (bs-0566R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
U-2OS cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (MADH7/Smad7) polyclonal Antibody, Unconjugated (bs-0566R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Sample:
Lane 1: Stomach (Mouse) Lysate at 40 ug
Lane 2: Spleen (Mouse) Lysate at 40 ug
Lane 3: Lung (Mouse) Lysate at 40 ug
Primary: Anti-MADH7/Smad7 (bs-0566R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 50 kD
Observed band size: 50 kD
Lane 1: Stomach (Mouse) Lysate at 40 ug
Lane 2: Spleen (Mouse) Lysate at 40 ug
Lane 3: Lung (Mouse) Lysate at 40 ug
Primary: Anti-MADH7/Smad7 (bs-0566R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 50 kD
Observed band size: 50 kD
Paraformaldehyde-fixed, paraffin embedded (rat kidney); 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 (MADH7) Polyclonal Antibody, Unconjugated (bs-0566R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse kidney); 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 (MADH7) Polyclonal Antibody, Unconjugated (bs-0566R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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SMAD7 Rabbit pAb(bs-0566R)-50ul/100ul/200ul
￥1180 - 2800
