Smad4 Antibody (YA4241)

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

Smad4 plays a central role in muscle physiology by regulating the balance between atrophy and hypertrophy. When recruited by MSTN, it promotes atrophy via phosphorylated SMAD2/4, while decreased MSTN leads to Smad4 release and subsequent recruitment by the BMP pathway to induce hypertrophy through phosphorylated SMAD1/5/8. It acts synergistically with SMAD1 and YY1 in BMP-mediated cardiac-specific gene expression and binds to SMAD binding elements (SBEs, 5'-GTCT/AGAC-3') within the BMP response element (BMPRE) of cardiac activating regions (By similarity). As a common SMAD (co-SMAD), Smad4 serves as a coactivator and mediator of TGF-beta signaling. It is a component of the heterotrimeric SMAD2/SMAD3-SMAD4 complex required for TGF-mediated signaling. Smad4 facilitates DNA binding of the SMAD2/SMAD4/FAST-1 complex and provides transcriptional activation function for SMAD1 or SMAD2. It also participates in the multimeric SMAD3/SMAD4/JUN/FOS complex at the AP1 promoter site, enabling synergistic transcriptional activity in response to TGF-beta. Smad4 may act as a tumor suppressor and positively regulates PDPK1 kinase activity by promoting its dissociation from the inhibitory 14-3-3 protein YWHAQ.

Q13485

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

affinity purified.

Non-conjugated

Primary Antibody; Mouse 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.
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  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

Liquid

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.