WB result of SMAD3 Recombinant Rabbit mAb
Primary antibody: SMAD3 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HT-1080 whole cell lysate 20 µg
Lane 2: HeLa whole cell lysate 20 µg
Lane 3: Jurkat whole cell lysate 20 µg
Lane 4: K562 whole cell lysate 20 µg
Lane 5: A431 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 48 kDa
Observed MW: 55 kDa
SMAD3 Recombinant Rabbit mAb (S-3147-36)
SMAD3 Recombinant Rabbit mAb (S-3147-36)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | SMAD3 |
| Synonyms | Mothers against decapentaplegic homolog 3; MAD homolog 3; Mad3; Mothers against DPP homolog 3; hMAD-3; JV15-2; SMAD family member 3 (SMAD 3; Smad3; hSMAD3); MADH3 |
| Immunogen | Recombinant Protein |
| Location | Cytoplasm, Nucleus |
| Accession | P84022 |
| Clone Number | S-3147-36 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC |
| Reactivity | Hu, Ms, Rt, Mk |
| Positive Sample | HT-1080, HeLa, Jurkat, K562, A431, RAW264.7, C6, COS-7 |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu, Ms, Rt, Mk |
| IHC-P | 1:1000 | Hu, Ms |
| ICC | 1:500 | Hu |
Background
SMAD3 protein is a key intracellular signaling transducer in the Transforming Growth Factor-β (TGF-β) signaling pathway. As one of the receptor-regulated SMADs (R-SMADs), SMAD3 is phosphorylated and activated upon binding of TGF-β superfamily ligands (such as TGF-β and activin) to their cell membrane receptors. It then forms a complex with SMAD4 and translocates into the nucleus, where it functions as a transcription factor to directly regulate the expression of target genes. It is extensively involved in physiological processes such as cell proliferation, differentiation, apoptosis, extracellular matrix formation, and immune regulation. SMAD3 plays a dual role in various pathological conditions, including fibrosis, cancer, and inflammatory diseases (it can suppress early tumor growth but also promote late-stage tumor progression and metastasis). Its activity is finely regulated by post-translational modifications such as ubiquitination and acetylation, as well as by inhibitory SMADs (e.g., SMAD7), making it an important potential therapeutic target for diseases.
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Picture
Western Blot
WB result of SMAD3 Recombinant Rabbit mAb
Primary antibody: SMAD3 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: RAW264.7 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 48 kDa
Observed MW: 552 kDa
WB result of SMAD3 Recombinant Rabbit mAb
Primary antibody: SMAD3 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: C6 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 48 kDa
Observed MW: 55 kDa
WB result of SMAD3 Recombinant Rabbit mAb
Primary antibody: SMAD3 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: COS-7 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 48 kDa
Observed MW: 55 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human colon. Anti-SMAD3 antibody was used at 1/1000 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 cervical squamous cell carcinoma. Anti-SMAD3 antibody was used at 1/1000 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 breast cancer. Anti-SMAD3 antibody was used at 1/1000 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 mouse kidney. Anti-SMAD3 antibody was used at 1/1000 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.
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti- Smad3 antibody was used at 1/500 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 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
