SMAD3 phospho S423/phospho S425 Antibody
Anti-Smad3 pS423 pS425 Antibody - Immunohistochemistry
Rabbit anti-SMAD pS423 pS425
Anti-Smad3 pS423 pS425 Antibody - Western Blot
Western Blot of Anti-SMAD3 pS423 pS425
Western Blot of Anti-SMAD3 pS423 pS425
Western Blot of Anti-SMAD3 pS423 pS425
Western Blot of Anti-SMAD3 pS423 pS425
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Datasheet

SMAD3 phospho S423/phospho S425 Antibody

Rabbit Polyclonal

600-401-919S 600-401-919
25 µL 100 µg
ELISA, IHC, WB, IF, IP
Human
Rabbit
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$99.00 /Per Item
$399.00 /Per Item
25 µL $99.00
100 µg $399.00
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Description

Background

This antibody is designed, produced, and validated as part of a collaboration between Rockland and the National Cancer Institute (NCI) and is suitable for Cancer, Immunology and Nuclear Signaling research. Smad3 (also known as Mothers against decapentaplegic homolog 3 Mothers against DPP homolog 3, Mad3, hMAD-3, JV15-2 or hSMAD3) is a transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinase.   These activators exert diverse effects on a wide array of cellular processes. The Smad proteins mediate much of the signaling responses induced by the TGF-b superfamily.  Briefly, activated type I receptor phosphorylates receptor-activated Smads (R-Smads) at their c-terminal two extreme serines in the SSXS motif, e.g. Smad2 and Smad3 proteins in the TGF-b pathway, or Smad1, Smad5 or Smad8 in the BMP pathway.  Then the phosphorylated R-Smad translocated into nucleus, where they regulate transcription of target genes.  Based on microarray and animal model experiments, Smad3 accounts for at least 80% of all TGF-b-mediated response.

Application Note

This affinity purified antibody has been tested for use in ELISA, immunohistochemistry, and western blot.  Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 48 kDa in size corresponding to phosphorylated Smad3 protein by western blotting in the appropriate stimulated tissue or cell lysate or extract.  Less than 0.2% reactivity is observed against the non-phosphorylated form of the immunizing peptide.  This antibody is phospho specific for dual phosphorylated pS423 and pS425 of Smad3. Stimulation with 2 ng/ml TGF-beta for 1 hour is suggested.

Purity/Specificity

This affinity-purified antibody is directed against the phosphorylated form of human Smad3 protein at the pS423 and pS425 residues. The product was affinity purified from monospecific antiserum by immunoaffinity purification.  Antiserum was first purified against the phosphorylated form of the immunizing peptide.  The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide.  Reactivity occurs against human Smad3 pS423 and pS425 protein and the antibody is specific for the phosphorylated form of the protein.   Reactivity with non-phosphorylated human Smad3 is minimal by ELISA and western blot.  Expect reactivity against phosphorylated Smad1 and Smad5.  Negligible reactivity is seen against other phosphorylated Smad family members.  A BLAST analysis was used to suggest cross reactivity with Smad3 from human, Xenopus laevis, Xenopus tropicalis, zebrafish, rat, mouse, swine, bovine and chicken based on 100% sequence homology with the immunogen.  Reactivity against homologues from other sources is not known.

Anti-SMAD3 pS423 pS425 (RABBIT) Antibody - 600-401-919
rabbit anti-SMAD3 pS423pS425 antibody, SMAD-3, SMAD 3, mothers against decapentaplegic homolog 3 antibody, MAD homolog 3, Mothers against DPP homolog 3, SMAD family member 3, MADH3, MADH 3, JV15-2, nothing
SMAD3
ELISA, IHC, WB
IF, IP
Human
Human
Phosphorylation
Rabbit
Polyclonal
IgG
Peptide
Anti-SMAD3 pS423pS425 antibody was prepared from whole rabbit serum produced by repeated immunizations with a dual phosphorylated synthetic peptide corresponding to a c-terminal region with Serine 423 and Serine 425 of human SMAD3 protein.
Liquid (sterile filtered)
1.16 mg/ml by UV absorbance at 280 nm
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
0.01% (w/v) Sodium Azide
None
Dry Ice
Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Expiration date is one (1) year from date of opening.
Iida R et al. Deficiency of M-LP/Mpv17L leads to development of β-cell hyperplasia and improved glucose tolerance via activation of the Wnt and TGF-β pathways. Biochim Biophys Acta Mol Basis Dis. (2022)
Applications
Undefined
Wang W et al. SARS‐CoV‐2 N Protein Induces Acute Kidney Injury via Smad3‐Dependent G1 Cell Cycle Arrest Mechanism. Adv Sci (Weinh). (2022)
Applications
IF, Confocal Microscopy; IHC, ICC, Histology
Tang PCT et al. Smad3 Promotes Cancer‐Associated Fibroblasts Generation via Macrophage–Myofibroblast Transition. Adv Sci (Weinh). (2022)
Applications
WB, IB, PCA
Pinkaew D et al. Fortilin interacts with TGF-β1 and prevents TGF-β receptor activation. Commun Biol. (2022)
Applications
WB, IB, PCA
Bosch-Queralt M et al. Diet-dependent regulation of TGFβ impairs reparative innate immune responses after demyelination. Nat Metab. (2021)
Applications
Undefined
Tamura Y et al. Anti-pyroptotic function of TGF-β is suppressed by a synthetic dsRNA analogue in triple negative breast cancer cells. Mol Oncol. (2021)
Applications
IHC, ICC, Histology
Dong L et al. Deletion of Smad3 protects against diabetic myocardiopathy in db/db mice. J Cell Mol Med. (2021)
Applications
IHC, ICC, Histology
Takagaki Y et al. Endothelial autophagy deficiency induces IL6-dependent endothelial mesenchymal transition and organ fibrosis. Autophagy. (2020)
Applications
Undefined
Hutchinson LD et al. Salt-inducible kinases (SIKs) regulate TGFβ-mediated transcriptional and apoptotic responses. Cell Death Dis. (2020)
Applications
WB, IB, PCA
Kang H et al. Somatic SMAD3-activating mutations cause melorheostosis by up-regulating the TGF-β/SMAD pathway. J Exp Med. (2020)
Applications
WB, IB, PCA
Navarro R et al. TGF‐β‐induced IGFBP‐3 is a key paracrine factor from activated pericytes that promotes colorectal cancer cell migration and invasion. Mol Oncol. (2020)
Applications
WB, IB, PCA
Ni J et al. Dual deficiency of angiotensin‐converting enzyme‐2 and Mas receptor enhances angiotensin II‐induced hypertension and hypertensive nephropathy. J Cell Mol Med. (2020)
Applications
IHC, ICC, Histology
Kim IY et al. Deletion of Akt1 Promotes Kidney Fibrosis in a Murine Model of Unilateral Ureteral Obstruction. Biomed Res Int. (2020)
Applications
WB, IB, PCA
Yang F et al. Inhibition of dipeptidyl peptidase-4 accelerates epithelial–mesenchymal transition and breast cancer metastasis via the CXCL12/CXCR4/mTOR axis. Cancer Res. (2019)
Applications
IF, Confocal Microscopy; WB, IB, PCA
Gao R et al. βklotho is essential for the anti‐endothelial mesenchymal transition effects of N‐acetyl‐seryl‐aspartyl‐lysyl‐proline. FEBS Open Bio. (2019)
Applications
WB, IB, PCA
Wei X et al. Spatially restricted stromal Wnt signaling restrains prostate epithelial progenitor growth through direct and indirect mechanisms. Cell Stem Cell. (2019)
Applications
Undefined
Stappenbeck F et al. Inhibition of Non-Small Cell Lung Cancer Cells by Oxy210, an Oxysterol-Derivative that Antagonizes TGFβ and Hedgehog Signaling. Cells. (2019)
Applications
WB, IB, PCA
Feng et al. TGF-β Mediates Renal Fibrosis via the Smad3-Erbb4-IR Long Noncoding RNA Axis. Molecular Therapy (2018)
Applications
IHC, ICC, Histology
Li et al. Fatty acid receptor modulator PBI-4050 inhibits kidney fibrosis and improves glycemic control. JCI Insight (2018)
Applications
IHC, ICC, Histology; WB, IB, PCA
Tang et al. Generation of Smurf2 Conditional Knockout Mice. International Journal of Biological Sciences (2018)
Applications
WB, IB, PCA
Chung et al. TGF-β promotes fibrosis after severe acute kidney injury by enhancing renal macrophage infiltration. JCI Insight (2018)
Applications
IF, Confocal Microscopy; WB, IB, PCA
Huang H et al. Lethal (3) malignant brain tumor-like 2 (L3MBTL2) protein protects against kidney injury by inhibiting the DNA damage–p53–apoptosis pathway in renal tubular cells. Kidney Int. (2018)
Applications
Undefined
Liu L et al. Neuronal transforming growth factor beta signaling via SMAD3 contributes to pain in animal models of chronic pancreatitis. Gastroenterology. (2018)
Applications
IHC, ICC, Histology
tang et al. Transforming Growth Factor-β (TGF-β) Directly Activates the JAK1-STAT3 Axis to Induce Hepatic Fibrosis in Coordination with the SMAD Pathway. Journal of Biological Chemistry (2017)
Applications
WB, IB, PCA
Li et al. FGFR1 is critical for the anti-endothelial mesenchymal transition effect of N-acetyl-seryl-aspartyl-lysyl-proline via induction of the MAP4K4 pathway. Cell Death & Disease (2017)
Applications
WB, IB, PCA
Tripathi V, Sixt KM, Gao S, et al Direct Regulation of Alternative Splicing by SMAD3 through PCBP1 Is Essential to the Tumor-Promoting Role of TGF-β. Mol Cell. (2016)
Applications
IP, Co-IP; WB, IB, PCA
Luo et al. A novel profibrotic mechanism mediated by TGFβ-stimulated collagen prolyl hydroxylase expression in fibrotic lung mesenchymal cells. The Journal of Pathology (2015)
Applications
WB, IB, PCA
Subathra et al. Kidney glycosphingolipids are elevated early in diabetic nephropathy and mediate hypertrophy of mesangial cells. American Journal of Physiology Renal Physiology (2015)
Applications
WB, IB, PCA
Herhaus L. et al. OTUB1 enhances TGFβ signalling by inhibiting the ubiquitylation and degradation of active SMAD2/3. Nat Commun. (2013)
Applications
WB, IB, PCA
Kawamura I et al. SnoN suppresses maturation of chondrocytes by mediating signal cross-talk between transforming growth factor-β and bone morphogenetic protein pathways. J Biol Chem. (2012)
Applications
IHC, ICC, Histology
Coffman JA et al. Oral–aboral axis specification in the sea urchin embryo: III. Role of mitochondrial redox signaling via H2O2. Dev Biol. (2009)
Applications
IF, Confocal Microscopy
Yamashita M. et al. TRAF6 mediates Smad-independent activation of JNK and p38 by TGF-beta. Mol Cell. (2008)
Applications
IP, Co-IP; WB, IB, PCA

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This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.

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