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宿主来源
Rabbit抗原名称
Smad4分子别名
Mothers against decapentaplegic homolog 4, MAD homolog 4, Mothers against DPP homolog 4, Deletion target in pancreatic carcinoma 4, SMAD 4, Smad4, hSMAD4免疫原
Synthetic Peptide细胞定位
CytoplasmAccession
Q13485克隆号
SDT-168-62抗体类型
Recombinant mAb反应种属 ?
Hu纯化方式
Protein A浓度
1 mg/ml标记
Unconjugated性状
Liquid缓冲体系
PBS储存条件
12 months from date of receipt, 4°C as supplied
应用
IHC-P ?
FCM
ICC ?
WB
稀释度
应用 稀释度 WB 1:1000 FCM 1:500 ICC 1:100 IHC-P 1:100
SMAD (mothers against decapentaplegic homologs) molecules are the core components in TGF-β signaling pathway. TGF-β binding to its receptor induces phosphorylation and activation of receptor-regulated SMADs (R-SMADs), SMAD2 and SMAD3, which subsequently associate with their partner SMAD4 and translocate from cytoplasm to nucleus. Formation of R-SMAD–SMAD4 complexes is essential in signaling of most TGF-β family members.
免疫印迹
WB result of SMAD4 Rabbit mAb Primary antibody: SMAD4 Rabbit mAb at 1/1000 dilution Lane 1: HT-29 whole cell lysate 20 µg Lane 2: HepG2 whole cell lysate 20 µg Lane 3: HCT 116 whole cell lysate 20 µg Lane 4: Jurkat whole cell lysate 20 µg Negative control: HT-29 whole cell lysate Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 60 kDa Observed MW: 65 kDa
流式分析
Flow cytometric analysis of HT-29 (Human colorectal adenocarcinoma epithelial cell, left) / HepG2 (Human hepatocellular carcinoma epithelial cell, right) cells labelling SMAD4 antibody at 1/500 dilution (0.1 μg)/ (red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody. Negative control: HT-29 cells
免疫组化
IHC shows positive staining in paraffin-embedded human pancreatic cancer (Loss of SMAD4 expression in tumors but expression in stromal and paracancerous cells). Anti-SMAD4 antibody was used at 1/100 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human pancreatic cancer (Loss of SMAD4 expression in tumors but nuclear expression in stromal cells). Anti-SMAD4 antibody was used at 1/100 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human lung squamous cell carcinoma. Anti-SMAD4 antibody was used at 1/100 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti-SMAD4 antibody was used at 1/100 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti-SMAD4 antibody was used at 1/100 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
免疫细胞化学
Negative control: ICC shows negative staining in HT-29 cells. Anti-SMAD4 antibody was used at 1/100 dilution and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue).
ICC shows positive staining in HepG2 cells. Anti-SMAD4 antibody was used at 1/100 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue).
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