Smad4 抗体 (YA4241)

Smad4 Antibody (YA4241) is a Mouse-derived and non-conjugated IgG1 monoclonal antibody, targeting to Smad4.

Smad4 在肌肉生理学中扮演着关键角色，调节肌肉萎缩 (atrophy) 和肥大 (hypertrophy) 的平衡。当被 MSTN 招募时，Smad4 通过磷酸化的 SMAD2/4 促进萎缩反应；而 MSTN 减少会导致 Smad4 释放，随后被 BMP 通路招募，通过磷酸化的 SMAD1/5/8 促进肥大。它与 SMAD1 和 YY1 协同作用，参与骨形态发生蛋白 (BMP) 介导的心脏特异性基因表达，并能结合心脏激活区域 BMP 反应元件 (BMPRE) 中的 SMAD 结合元件 (SBEs，5'-GTCT/AGAC-3') (By similarity)。作为通用 SMAD (co-SMAD)，Smad4 是 TGF-β (转化生长因子) 信号转导的共激活因子和媒介。它是异源三聚体 SMAD2/SMAD3-SMAD4 复合物的组成部分，该复合物在细胞核中形成，是 TGF 介导的信号传导所必需的。此外，Smad4 促进 SMAD2/SMAD4/FAST-1 复合物与 DNA 的结合，并为 SMAD1 或 SMAD2 提供激活转录的功能。它还参与形成多聚体 SMAD3/SMAD4/JUN/FOS 复合物，该复合物在 AP1 启动子位点形成，响应 TGF-β 具有协同转录活性。Smad4 可能具有肿瘤抑制功能，并通过刺激 PDPK1 激酶与负调控因子 14-3-3 蛋白 YWHAQ 解离来正向调节 PDPK1 激酶活性。

Q13485

Predicted band size: 60 kDa; Observed band size: 65 kDa

affinity purified.

Non-conjugated

Primary Antibody; Mouse Monoclonal Antibody

Monoclonal Antibody

Mouse

Human, Mouse

WB: 1:500-1:2000; IHC: 1:200-1:1000; IF: 1:200-1:1000; FC: 1:200-1:400; ELISA: 1:10000

• 
  Western blot analysis of extracts from A431(lane 2(20μg), K562 (lane 3(20μg) and HepG2 (lane 4(20μg) using SMAD4 (30091) Mouse mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of NIH/3T3 cells labeling SMAD4 with SMAD4 Antibody (HY-P84544)at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with SMAD4 Antibody (HY-P84544) at 1/200 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Mouse IgG H&L（HY-P8005, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of NIH/3T3 cells labeling SMAD4 with SMAD4 Antibody (HY-P84544) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with SMAD4 Antibody (HY-P84544) at 1/500 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Mouse IgG H&L（HY-P8005, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded Mouse lung tissue using SMAD4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (30091, 1/500) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded Mouse lung tissue using SMAD4 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (30091, 1/500) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, IHC-P, ICC/IF, FC, ELISA

液体

Supplied in PBS with 0.05% sodium azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG1

Purified recombinant fragment of human SMAD4 aa 336-552.