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Purified anti-SMAD4 Antibody

Pricing & Availability
Clone
W23084C (See other available formats)
Regulatory Status
RUO
Other Names
Mothers against decapentaplegic homolog 4, MAD homolog 4, Mothers against DPP homolog 4, Deletion target in pancreatic carcinoma 4, DPC4, MADH4
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
a. W23084C_PURE_SMAD4_1_100125
Whole cell extracts (15 µg total protein per lane) from indicated cell lines (HAP1 SMAD4 KO, Horizon Cat. No. HZGHC005409c003) and were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-SMAD4 (clone W23084C) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • a. W23084C_PURE_SMAD4_1_100125
    Whole cell extracts (15 µg total protein per lane) from indicated cell lines (HAP1 SMAD4 KO, Horizon Cat. No. HZGHC005409c003) and were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-SMAD4 (clone W23084C) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • b. W23084C_PURE_SMAD4_2_100125
    Whole cell extracts (15 µg total protein per lane) from indicated cell lines and were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-SMAD4 (clone W23084C) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-GAPDH (Cat. No. 607903) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • c. W23084C_PURE_SMAD4_3_100125
    IHC staining with Purified anti-SMAD4 (clone W23084C) on formalin-fixed paraffin-embedded human placenta tissue. Following antigen retrieval using 1X Tris-EDTA pH 9.0 Antigen Retrieval Buffer (Cat. No. 422704), the tissue was incubated with (panel A) and without (panel B) Purified anti-SMAD4 (clone W23084C) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 40X objective and merged (panels C and D). Scale bar: 50 µm
  • d. W23084C_PURE_SMAD4_4_100125
    IHC staining with Purified anti-SMAD4 (clone W23084C) on formalin-fixed paraffin-embedded human colon tissue. Following antigen retrieval using 1X Tris-EDTA pH 9.0 Antigen Retrieval Buffer (Cat. No. 422704), the tissue was incubated with (panel A) and without (panel B). Purified anti-SMAD4 (clone W23084C) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 40X objective and merged (panels C and D). Scale bar: 50 µm
  • e. W23084C_PURE_SMAD4_5_100125
    HAP1 wild-type cells (positive cell line, filled histogram) and HAP1 SMAD4 Knockout cells (negative cell line, open histogram) (Horizon Cat. No. HZGHC005409c003) were fixed and permeabilized using True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401) and intracellularly stained with Purified anti-SMAD4 (clone W23084C), or Purified Rat IgG2a, κ Isotype Control (dashed histogram, representative of both the positive and negative cell lines) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416).
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648884 25 µg 124€
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648885 100 µg 340€
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Description

SMAD4, also known as Deleted in Pancreatic Carcinoma locus 4 (DPC4), is a pivotal mediator of transforming growth factor-β (TGF-β) superfamily signaling. SMAD4 belongs to the common-mediator SMAD (co-SMAD) subclass and functions as a necessary partner for receptor-regulated SMADs (R-SMADs) in the canonical TGF-β, activin, and bone morphogenetic protein (BMP) pathways. 
In the canonical TGF-β superfamily cascade, ligand binding induces heteromeric complex formation between type II and type I serine/threonine kinase receptors, leading to phosphorylation of R-SMADs. Phosphorylated R-SMADs associate with SMAD4 to form hetero-oligomeric complexes that translocate into the nucleus. These complexes bind SMAD-binding elements in target gene promoters and recruit transcriptional co-activators or co-repressors, modulating the expression of genes governing cell cycle arrest, differentiation programs, apoptosis, and extracellular matrix remodeling. SMAD4 serves as a structural and functional scaffold essential for transcriptional regulation. Beyond canonical signaling, SMAD4 engages in extensive crosstalk with MAPK, PI3K–Akt, and Wnt/β-catenin pathways, enabling context-dependent modulation of transcriptional output.
SMAD4 dysfunction is implicated in diverse human diseases. SMAD4 is frequently inactivated in pancreatic ductal adenocarcinoma through deletion or inactivating mutation. Germline SMAD4 mutations cause hereditary syndromes such as Juvenile Polyposis Syndrome (JPS) and are characterized by hamartomatous gastrointestinal polyps with elevated risk of colorectal and gastric cancer. Gain-of-function mutations in SMAD4 underlie Myhre syndrome, featuring connective tissue overgrowth, short stature, craniofacial anomalies, and cardiovascular defects. SMAD4 also influences immune homeostasis by modulating T cell differentiation, particularly regulatory T cell development.

Product Details
Quality Statement

This product was carefully developed for performance and specificity using the following products from Horizon Discovery, a Revvity company: HAP1 SMAD KO cells (Exon 3) (Horizon Cat. No. HZGHC005409c003)

Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Recombinant fragment of human SMAD4
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
IHC-P, ICFC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.25 - 1.0 µg/mL. For immunohistochemistry, a concentration range of 2.5 - 10.0 µg/mL is suggested. For flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Additional Product Notes

For use in intracellular flow cytometry (ICFC), it is recommended to fix/permeabilize with True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401).

Fixation/permeabilization with True-Phos™ Perm Buffer (Cat. No. 425401) or Cyto-Fast™ Fix/Perm Buffer Set (Cat. No. 426803) is not recommended.

For use in immunohistochemistry on formalin-fixed paraffin-embedded tissue (IHC-P), it is recommended to perform antigen retrieval using either Citrate Buffer, 10X (Cat. No. 420902) or Tris-EDTA pH 9.0 Antigen Retrieval Buffer (10X) (Cat. No. 422704).

Antigen Details

Structure
SMAD4 is a 552 amino acid protein with a predicted molecular weight of 60.4 kD.
Distribution

Cytoplasm and nucleus

Function
Belongs to common SMAD (co-SMAD) and serves as the coactivator and mediator of signal transduction by TGF-beta.
Interaction
Forms heterotrimeric complex with SMAD2/SMAD3
Biology Area
Cancer Biomarkers, Cardiovascular Biology, Cell Biology, Cell Proliferation and Viability, Inhibitory Molecules, Signal Transduction
Molecular Family
Phospho-Proteins
Antigen References
  1. Wilentz RE, et al. 2000. Cancer Res. 60(7):2002–2006.
  2. Tascilar M, et al. 2001. Clin. Cancer Res. 7(12):4115–4121.
  3. Gallione CJ, et al. 2004. Lancet. 363(9412):852–859.
  4. Le Goff C, et al. 2012. Nat. Genet. 44(1):85–88. PMID:22158539.
Gene ID
4089 View all products for this Gene ID
UniProt
View information about SMAD4 on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 10/02/2025

For Research Use Only. Not for diagnostic or therapeutic use.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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