Rabbit Anti-Phospho-Smad2 (Thr220) antibody |
反应物种(预测) |
Mouse,Rat,Chicken,Dog,Cow,Horse |
产品应用(已验证) |
FCM |
产品应用(可尝试) |
WB,IHC,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=2ug/Test,IF=1:100-500, |
研究领域 |
肿瘤,免疫学,信号转导,转录调节因子 |
标签 |
Array |
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Tissue/cell: human placenta 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-Phospho-Smad2(Thr220) Polyclonal Antibody, Unconjugated(bs-5618R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
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Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-Phospho-Smad2 (Thr220) antibody (bs-5618R)
Dilution: 2μg /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.
RRID:AB_11070487
产品名称:Rabbit Anti-Phospho-Smad2 (Thr220) antibody
别名: Smad2 (phospho T220); p-Smad2 (phospho T220); phospho-Smad2(p-Thr220); p-Smad2(Thr220); phospho-Smad2(p-Thr220); hMAD 2; hSMAD2; JV18 1; JV18; JV181; MAD; MAD Related Protein 2; MADH2; MADR2; MGC22139; MGC34440; Mothers Against Decapentaplegic Homolog 2;
中文名称:磷酸化细胞信号转导分子SMAD2抗体
英文名称:Rabbit Anti-Phospho-Smad2 (Thr220) antibody
抗体来源: Rabbit
克隆类型:多克隆
细胞定位:细胞核,细胞浆
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 Smad2 around the phosphorylation site of Thr220
抗原表位:PE(p-T)PP
SWISS:Q15796
Gene ID :4087
Human Gene ID:4087
Smad2 is a 58 kDa member of a family of proteins involved in cell proliferation, differentiation and development. The Smad family is divided into three subclasses: receptor-regulated Smad's, activin/TGF alpha receptor-regulated (Smad2 and 3) or BMP receptor regulated (Smad1, 5, and 8); the common partner, (Smad4) that functions via its interaction to the various Smad's; and the inhibitory Smad's, (Smad6 and Smad7). Smad2 consists of two highly conserved domains, the N terminal Mad homology (MH1) and the C-terminal Mad homology 2 (MH2) domains. The MH1 domain binds DNA and regulates nuclear import and transcription while the MH2 domain conserved among all the Smad's regulates Smad2 oligomerization and binding to cytoplasmic adaptors and transcription factors. Activated Smad2 associates with Smad4 and translocates as a complex into the nucleus, allowing its binding to DNA and transcription factors. This translocation of Smad2 (as well as Smad3) into the nucleus is a central event in TGF beta signaling. Phosphorylation of threonine 8 in the calmodulin binding region of the MH1 domain by extracellular signal regulated kinase 1(ERK 1) enhances Smad2 transcriptional activity, which is negatively regulated by calmodulin. The regulation of Smad2 phosphorylation on threonine 8 by ERK 1 and calmodulin is critical for Smad2 mediated 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:Momomer; the absence of TGF-beta. Heterodimer; in the presence of TGF-beta. Forms a heterodimer with co-SMAD, SMAD4, in the nucleus to form the transactivation complex SMAD2/SMAD4. Interacts with AIP1, HGS, PML and WWP1. Interacts with NEDD4L in response
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. On dephosphorylation by phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of
Tissue Specificity:Expressed at high levels in skeletal muscle, heart and placenta.
Post-translational modifications:Phosphorylated on one or several of Thr-220, Ser-245, Ser-250, and Ser-255. In response to TGF-beta, phosphorylated on Ser-465/467 by TGF-beta and activin type 1 receptor kinases. Able to interact with SMURF2 when phosphorylated on Ser-465/467, recruiting
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.