| Rabbit Anti-Phospho-Smad3 (Ser425) antibody |
| 反应物种（预测） |
Human,Chicken,Pig,Cow,Horse,Rabbit |
| 产品应用（已验证） |
WB,IHC,FCM |
| 产品应用（可尝试） |
ICC,IF,ELISA |
| 推荐稀释比例 |
WB=1:500-2000,Elisa=1:5000-10000,IHC-P=1:100-500,IHC-F=1:100-500,Flow Cyt=1μg/Test,IF=1:100-500,ICC=1:100-500, |
| 研究领域 |
肿瘤,细胞生物,免疫学,信号转导,干细胞,细胞凋亡,生长因子和激素,转录调节因子 |
| 标签 |
Array |
-
Tissue/cell: human colon carcinoma; 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-Phospho-Smad3(Ser425) Polyclonal Antibody, Unconjugated(bs-5616R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
-
Paraformaldehyde-fixed, paraffin embedded (Human colon cancer); 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-Smad3 (Ser425)) Polyclonal Antibody, Unconjugated (bs-5616R) at 1:400 overnight at 4°C, followed by a conjugated secondary antibody (sp-0023) for 20 minutes and DAB staining.
-
Blank control (Black line): HUVEC (Black).
Primary Antibody (green line): Rabbit Anti-Phospho-Smad3 (Ser425) antibody (bs-5616R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): 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 room temperature. 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: Heart (Mouse) Lysate at 40 ug
Lane 3: Kidney (Mouse) Lysate at 40 ug
Lane 4: Cerebrum (Rat) Lysate at 40 ug
Primary: Anti- Phospho-Smad3 (Ser425) (bs-5616R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 52 kD
Observed band size: 50 kD
-
Paraformaldehyde-fixed, paraffin embedded (mouse brain); 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-Smad3 (Ser425)) Polyclonal Antibody, Unconjugated (bs-5616R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
-
Paraformaldehyde-fixed, paraffin embedded (rat brain); 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-Smad3 (Ser425)) Polyclonal Antibody, Unconjugated (bs-5616R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
RRID：AB_11109578
产品名称：Rabbit Anti-Phospho-Smad3 (Ser425) antibody
别名： SMAD3(phospho S425); Smad3 (phospho Ser425); p-Smad3 (Ser425); hMAD 3; hSMAD3; HSPC193; JV15 2; JV152; MAD (mothers against decapentaplegic Drosophila) homolog 3; MAD3; MADH 3; MADH3; Mothers against decapentaplegic homolog 3; Mothers against DPP homolog
中文名称：磷酸化细胞信号转导分子SMAD3抗体
英文名称：Rabbit Anti-Phospho-Smad3 (Ser425) antibody
抗体来源： Rabbit
克隆类型：多克隆
细胞定位：细胞核,细胞浆,细胞膜
性 状：Liquid
亚 型：IgG
纯化方法：affinity purified by Protein A
保存条件：Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
免 疫 原：KLH conjugated Synthesised phosphopeptide derived from human Smad3 around the phosphorylation site of Ser425
抗原表位：SV(p-S)-NH2
SWISS：P84022
Gene ID ：4088
Human Gene ID：4088
Smad3 is a 50 kDa member of a family of proteins that act as key mediators of TGF beta superfamily signaling in cell proliferation, differentiation and development. The Smad family is divided into three subclasses: receptor regulated Smads, activin/TGF beta receptor regulated (Smad2 and 3) or BMP receptor regulated (Smad 1, 5, and 8); the common partner, (Smad4) that functions via its interaction to the various Smads; and the inhibitory Smads, (Smad6 and 7). Activated Smad3 oligomerizes with Smad4 upon TGF beta stimulation and translocates as a complex into the nucleus, allowing its binding to DNA and transcription factors. Phosphorylation of the two TGF beta dependent serines 423 and 425 in the C terminus of Smad3 is critical for Smad3 transcriptional activity and TGF beta signaling.
Function：Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes th
Subunit：Monomer; in the absence of TGF-beta. Homooligomer; in the presence of TGF-beta. Heterotrimer; forms a heterotrimer in the presence of TGF-beta consisting of two molecules of C-terminally phosphorylated SMAD2 or SMAD3 and one of SMAD4 to form the transcrip
Subcellular Location：Cytoplasm. Nucleus. Note=Cytoplasmic and nuclear in the absence of TGF-beta. On TGF-beta stimulation, migrates to the nucleus when complexed with SMAD4. Through the action of the phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out o
Post-translational modifications：Phosphorylated on serine and threonine residues. Enhanced phosphorylation in the linker region on Thr-179, Ser-204 and Ser-208 on EGF AND TGF-beta treatment. Ser-208 is the main site of MAPK-mediated phosphorylation. CDK-mediated phosphorylation occurs in
DISEASE：Defects in SMAD3 may be a cause of colorectal cancer (CRC) [MIM:114500].
Defects in SMAD3 are the cause of Loeys-Dietz syndrome type 1C (LDS1C) [MIM:613795]. LDS1C is an aortic aneurysm syndrome with widespread systemic involvement. The disorder is ch
Similarity：Belongs to the dwarfin/SMAD family.
Contains 1 MH1 (MAD homology 1) domain.
Contains 1 MH2 (MAD homology 2) domain.
Important Note：This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
400-901-9800
说明书
联系我们
打印此页面
收藏