Phospho-SMAD2 (Ser250) Antibody

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Description

Western Blot Studies

  • TGF-β signaling: The antibody successfully detects phosphorylation of SMAD2 at Ser250 in human 293T cells treated with PMA/TPA or Calyculin A , confirming its role in TGF-β-induced activation.

  • Cancer research: In MCF-7 breast cancer cells, the antibody identified phosphorylated SMAD2 in response to TGF-β stimulation, correlating with increased transcriptional activity .

Immunofluorescence

  • Subcellular localization: The antibody has been used to visualize nuclear translocation of phosphorylated SMAD2 in cells treated with TGF-β, highlighting its role in transcriptional regulation .

Cross-Reactivity and Specificity

  • Species compatibility: Predicted reactivity with zebrafish, bovine, and sheep based on sequence alignment , making it suitable for comparative studies across species.

  • Epitope specificity: The antibody targets the phosphorylated Ser250 residue, part of the MH2 domain critical for SMAD2/SMAD4 complex formation .

Role in TGF-β Signaling Research

Phosphorylation at Ser250 is a hallmark of SMAD2 activation by TGF-β type I receptor kinases. This modification facilitates:

  • Complex formation: Phosphorylated SMAD2 binds SMAD4, enabling nuclear translocation .

  • Transcriptional regulation: The complex activates genes involved in cell cycle arrest, apoptosis, and extracellular matrix production .

  • Pathological implications: Dysregulation of Ser250 phosphorylation is linked to cancers (e.g., colorectal carcinoma) and fibrotic diseases .

Experimental Protocols

  • Western Blot:

    • Lysis buffer: RIPA with phosphatase inhibitors .

    • Blocking: 3% BSA in TBST .

    • Primary dilution: 1:500–1:1000 overnight at 4°C .

  • Immunofluorescence:

    • Fixation: 4% paraformaldehyde .

    • Permeabilization: 0.1% Triton X-100 .

Product Specs

Form
Rabbit IgG in phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Lead Time
Typically, we can ship the products within 1-3 business days after receiving your orders. Delivery time may vary depending on the purchasing method or location. For specific delivery times, please consult your local distributors.
Synonyms
Drosophila, homolog of, MADR2 antibody; hMAD-2 antibody; HsMAD2 antibody; JV18 antibody; JV18-1 antibody; JV181 antibody; MAD antibody; MAD homolog 2 antibody; MAD Related Protein 2 antibody; Mad-related protein 2 antibody; MADH2 antibody; MADR2 antibody; MGC22139 antibody; MGC34440 antibody; Mother against DPP homolog 2 antibody; Mothers against decapentaplegic homolog 2 antibody; Mothers against decapentaplegic, Drosophila, homolog of, 2 antibody; Mothers against DPP homolog 2 antibody; OTTHUMP00000163489 antibody; Sma and Mad related protein 2 antibody; Sma- and Mad-related protein 2 MAD antibody; SMAD 2 antibody; SMAD family member 2 antibody; SMAD, mothers against DPP homolog 2 antibody; SMAD2 antibody; SMAD2_HUMAN antibody
Target Names
Uniprot No.

Target Background

Function
Phospho-SMAD2 (Ser250) Antibody targets the phosphorylated form of SMAD2, a receptor-regulated SMAD (R-SMAD) that plays a crucial role as an intracellular signal transducer and transcriptional modulator. This protein is activated by transforming growth factor-beta (TGF-beta) and activin type 1 receptor kinases. It binds to the TGF-beta responsive element (TRE) in the promoter region of numerous genes regulated by TGF-beta, and upon formation of the SMAD2/SMAD4 complex, initiates gene transcription. SMAD2 has been implicated as a potential tumor suppressor in colorectal carcinoma. Notably, it positively regulates PDPK1 kinase activity by promoting its dissociation from the 14-3-3 protein YWHAQ, which acts as a negative regulator.
Gene References Into Functions
  1. These findings suggest a more prominent role for SMAD3 and SMAD4 compared to SMAD2 in TGFbeta-induced chondrogenesis of human bone marrow-derived mesenchymal stem cells. PMID: 28240243
  2. The study revealed that miR4865p is upregulated in Osteoarthritis and may inhibit chondrocyte proliferation and migration by suppressing SMAD2. PMID: 29749497
  3. This research highlights the significance of the identified Sirt1-Smad2 interaction for the regulation of TGFbeta-dependent gene transcription. PMID: 29187201
  4. The current study indicated that S100A11 promotes EMT through an increase in TGF-beta1 expression, leading to TGF-beta1-induced upregulation of p-SMAD2 and 3. PMID: 29569474
  5. The findings suggest that miR2145p may promote the adipogenic differentiation of BMSCs through regulation of the TGFbeta/Smad2/COL4A1 signaling pathway, potentially leading to the development of a novel drug for postmenopausal osteoporosis. PMID: 29532880
  6. High SMAD2 expression is associated with fibrosis in chronic pancreatitis and pancreatic cancer. PMID: 29328490
  7. The results indicate that co-expression of active SMAD2/3 could enhance multiple types of transcription factor (TF)-based cell identity conversion, making it a powerful tool for cellular engineering. PMID: 29174331
  8. The study demonstrated that ITZ treatment effectively suppresses EMT, and this effect is partially mediated by impaired TGF-b/SMAD2/3 signaling. The role of TGF-b/SMAD2/3 signaling in mediating the effect of ITZ was further confirmed by the observation that recombinant TGF-b induced, but the TGF-b neutralizing antibody inhibited EMT as well as the invasion and migration of pancreatic cancer cells. PMID: 29484419
  9. The SMAD2/3 interactome reveals that TGFbeta controls m(6)A mRNA methylation in pluripotency. PMID: 29489750
  10. This study provides novel insights into how oscillatory shear stress regulates Smad2 signaling and pro-inflammatory genes through the complex signaling networks of integrins, transforming growth factor-beta receptors, and extracellular matrices, highlighting the molecular basis of regional pro-inflammatory activation within disturbed flow regions in the arterial tree. PMID: 29295709
  11. The findings demonstrated that thymoquinone suppressed the metastatic phenotype and reversed EMT of prostate cancer cells by negatively regulating the TGF-beta/Smad2/3 signaling pathway. This suggests that thymoquinone is a potential therapeutic agent against prostate cancer by targeting TGF-beta. PMID: 29039572
  12. MicroRNA-486-5p suppresses TGFB2-induced proliferation, invasion, and epithelial-mesenchymal transition of lens epithelial cells by targeting Smad2. PMID: 29229876
  13. Treatment with iPSC-CM markedly reduced the proliferation of TGF-beta1-exposed cells, and the activities of TGF-beta1, Smad-2, and Smad-3. These changes in the expression of these molecules were accompanied by a significant improvement in the lung structure of mice with PF. PMID: 29115383
  14. The study found expression of pSmad2/3 and Smad4 in different liver tissues, with up-regulated expression of both antibodies in chronic hepatitis C with higher stages of fibrosis and higher grades of activity. PMID: 29924446
  15. TGFbeta and IL1beta signaling interact at the SMAD2/3 level in human primary MSC. Down-stream TGFbeta target genes were repressed by IL1beta independent of C-terminal SMAD2 phosphorylation. The study demonstrates that SMAD2/3 linker modifications are required for this interplay and identified TAK1 as a crucial mediator of IL1beta-induced TGFbeta signal modulation. PMID: 28943409
  16. The studies provide a molecular mechanism by which UCHL5 mitigates TGFbeta-1 signaling by stabilizing Smad2/Smad3. These data indicate that UCHL5 may contribute to the pathogenesis of idiopathic pulmonary fibrosis and may be a potential therapeutic target. PMID: 27604640
  17. The study demonstrated that the downregulation of CLDN6 is regulated through promoter methylation by DNMT1, which depends on the SMAD2 pathway, and that CLDN6 is a key regulator in the SMAD2/DNMT1/CLDN6 pathway to inhibit EMT, migration, and invasion of breast cancer cells. PMID: 28867761
  18. High Expression of Smad2 is associated with liver cancer. PMID: 28415588
  19. While autocrine signaling activates Smad2/3 in differentiating extravillous trophoblasts, paracrine factors contribute to Smad phosphorylation in these cells. PMID: 28864007
  20. Kidney samples from patients with advanced stages of diabetic nephropathy showed elevated pSmad2 staining. PMID: 28805484
  21. Smad2 (and myostatin) were significantly up-regulated in the failing heart of female patients, but not male patients. PMID: 28465115
  22. Nodal signaling through the Smad2/3 pathway up-regulated Slug, Snail, and c-Myc to induce EMT, thereby promoting Vasculogenic mimicry (VM) formation. PMID: 27659524
  23. This study shows that EGF induces epithelial-mesenchymal transition through phospho-Smad2/3-Snail signaling pathway in breast cancer cells. PMID: 27829223
  24. Multiple myeloma cells adapted to long-term exposure to hypoxia exhibit stem cell characteristics with TGF-beta/Smad pathway activation. PMID: 29309790
  25. A novel heterozygous missense mutation (c.833C>T, p.A278V) in the SMAD2 gene was identified in a family with early onset aortic aneurysms. PMID: 28283438
  26. Data suggest that oncogenic Y-box binding protein 1 (YB-1) indirectly enhances transforming growth factor beta (TGFbeta) signaling cascades via Sma/Mad related protein 2 (Smad2) phospho-activation and may represent a promising factor for future diagnosis and therapy of breast cancer. PMID: 29187452
  27. Asiaticoside hindered the invasive growth of KFs by inhibiting the GDF-9/MAPK/Smad pathway. PMID: 28346732
  28. High Smad2 expression is associated with invasion and metastasis in pancreatic ductal adenocarcinoma. PMID: 26908446
  29. Data indicate that miR-206 inhibits neuropilin-1 (NRP1) and SMAD2 gene expression by directly binding to their 3'-UTRs. PMID: 27014911
  30. Results show that members of the Activin branch of the TGFbeta signaling pathway, namely Put and Smad2, are autonomously required for cell and tissue growth in the Drosophila larval salivary gland. PMID: 28123053
  31. CytoD modified MKL1, a coactivator of serum response factor (SRF) regulating CTGF induction, and promoted its nuclear localization. PMID: 27721022
  32. Cells expressing mutant huntingtin have a dysregulated transcriptional response to epidermal growth factor stimulation. PMID: 27988204
  33. CRT regulates TGF-beta1-induced-EMT through modulating Smad signaling. PMID: 28778674
  34. P311 is a novel TGFbeta1/Smad signaling-mediated regulator of transdifferentiation in epidermal stem cells during cutaneous wound healing. PMID: 27906099
  35. Human epidermal growth factor receptor 2 (HER-2) levels were correlated well with TSP50/p-Samd2/3 and TSP50/p27 expression status. Thus, these studies revealed a novel regulatory mechanism underlying TSP50-induced cell proliferation and provided a new favorable intervention target for the treatment of breast cancer. PMID: 28650473
  36. IL-17 can induce A549 alveolar epithelial cells to undergo epithelial-mesenchymal transition via the TGF-beta1 mediated Smad2/3 and ERK1/2 activation. PMID: 28873461
  37. A critical role for miR-503-3p in induction of breast cancer EMT was identified. PMID: 28161325
  38. Nuclear localization of Smad2 was reduced in TGFbeta-1-stimulated primary tubular epithelial cells. Changes in nuclear Smad2 correlated with a reduced expression of the pro-fibrotic factor CTGF. Transient downregulation of Smad2 interfered with TGFbeta-1-induced CTGF synthesis. PMID: 27155083
  39. Low SMAD2 expression is associated with progression of hepatic fibrosis. PMID: 28423499
  40. To study the translation between mouse model and patients, the signature of phosphorylated Sma- and Mad-related protein 2 (pSmad2), as a molecular marker of TGF-beta/activin activity, was evaluated in the kidneys of streptozotocin (STZ)-treated mice compared to that of type 1 diabetes (T1D) patients. PMID: 28064277
  41. SMAD2/SMAD3 signaling by bone morphogenetic proteins causes disproportionate induction of HAS2 expression and hyaluronan production in immortalized human granulosa cells. PMID: 26992562
  42. miR-27a contributed to cell proliferation and invasion by inhibiting TGF-beta-induced cell cycle arrest. These results suggest that miR-27a may function as an oncogene by regulating SMAD2 and SMAD4 in lung cancer. PMID: 28370334
  43. cPLA2alpha activates PI3K/AKT and inhibits Smad2/3 during epithelial-mesenchymal transition of hepatocellular carcinoma cells. PMID: 28649002
  44. Selective inhibition of SMAD3 or CCT6A efficiently suppresses TGF-beta-mediated metastasis. These findings provide a mechanism that directs TGF-beta signaling toward its prometastatic arm and may contribute to the development of therapeutic strategies targeting TGF-beta for non-small-cell lung carcinoma. PMID: 28375158
  45. In response to TGF-beta, RASSF1A is recruited to TGF-beta receptor I and targeted for degradation by the co-recruited E3 ubiquitin ligase ITCH. RASSF1A degradation is necessary to permit Hippo pathway effector YAP1 association with SMADs and subsequent nuclear translocation of receptor-activated SMAD2. PMID: 27292796
  46. Smad2 is a key scaffold, allowing RIN1 to act as a GTP exchange factor for MFN2-GTPase activation to promote mitochondrial ATP synthesis and suppress superoxide production during mitochondrial fusion. PMID: 27184078
  47. Ang down-regulate the expression of Col-I, alpha-SMA, and TGF-beta1/Smad2/3, subsequently inhibiting fibroblast-myofibroblast transition. PMID: 27543459
  48. The findings suggest a stronger chondrogenic potential of CD105(+) SMSCs compared to CD105(-) SMSCs. The study indicates that CD105 enhances chondrogenesis of SMSCs by regulating the TGF-beta/Smad2 signaling pathway, but not Smad1/5. This research provides a better understanding of CD105 with respect to chondrogenic differentiation. PMID: 27107692
  49. The findings show that TIEG1 is highly expressed in human keloids and that it directly binds and represses Smad7 promoter-mediated activation of TGF-beta/Smad2 signaling. PMID: 28108300
  50. High expression of SMAD2 is associated with colorectal carcinoma. PMID: 27959430
Database Links

HGNC: 6768

OMIM: 601366

KEGG: hsa:4087

STRING: 9606.ENSP00000262160

UniGene: Hs.12253

Protein Families
Dwarfin/SMAD family
Subcellular Location
Cytoplasm. Nucleus.
Tissue Specificity
Expressed at high levels in skeletal muscle, endothelial cells, heart and placenta.

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