SMAD3 antibody (pSer423, pSer425)

Catalog No. ABIN129675

Supplier Product No.: 600-401-919

Product Details anti-SMAD3 Antibody

Target: SMAD3 (SMAD, Mothers Against DPP Homolog 3 (SMAD3))

Binding Specificity: pSer423, pSer425

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This SMAD3 antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Immunohistochemistry (IHC), Flow Cytometry (FACS)

Supplier Product No.: 600-401-919

Supplier: Rockland

Purpose: SMAD3 phospho S423/phospho S425 Antibody

Cross-Reactivity (Details): This affinity-purified antibody is directed against the phosphorylated form of human Smad3 protein at the pS423 and pS425 residues.

Characteristics: Synonyms: 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

Purification: The product was affinity purified from monospecific antiserum by immunoaffinity purification.

Sterility: Sterile filtered

Immunogen:

Immunogen: 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.

Immunogen Type: Conjugated Peptide

Isotype: IgG

Application Details

Application Notes:

Flow Cytometry Dilution: User Optimized

Immunohistochemistry Dilution: 1:500

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.

Western Blot Dilution: 1:2,000 - 1:20,000

ELISA Dilution: 1:15,000 - 1:30,000

Other: User Optimized

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 1.0 mg/mL

Buffer:

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Stabilizer: None

Preservative: 0.01 % (w/v) Sodium Azide

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: 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.

Expiry Date: 12 months

References for anti-SMAD3 antibody (ABIN129675)

Hutchinson, Darling, Nicolaou, Gori, Squair, Cohen, Hill, Sapkota: "Salt-inducible kinases (SIKs) regulate TGFβ-mediated transcriptional and apoptotic responses." in: Cell death & disease, Vol. 11, Issue 1, pp. 49, (2020) (PubMed).

Stappenbeck, Wang, Tang, Zhang, Parhami: "Inhibition of Non-Small Cell Lung Cancer Cells by Oxy210, an Oxysterol-Derivative that Antagonizes TGFβ and Hedgehog Signaling." in: Cells, Vol. 8, Issue 10, (2020) (PubMed).

Feng, Tang, Huang, Sun, You, Xiao, Lv, Xu, Lan: "TGF-β Mediates Renal Fibrosis via the Smad3-Erbb4-IR Long Noncoding RNA Axis." in: Molecular therapy : the journal of the American Society of Gene Therapy, Vol. 26, Issue 1, pp. 148-161, (2019) (PubMed).

Subramanian, Kanzaki, Galloway, Schilling: "Mechanical force regulates tendon extracellular matrix organization and tenocyte morphogenesis through TGFbeta signaling." in: eLife, Vol. 7, (2019) (PubMed).

Gao, Kanasaki, Li, Kitada, Okazaki, Koya: "βklotho is essential for the anti-endothelial mesenchymal transition effects of N-acetyl-seryl-aspartyl-lysyl-proline." in: FEBS open bio, Vol. 9, Issue 5, pp. 1029-1038, (2019) (PubMed).

Tang, Tang, Xu, Li, Deng, Zhang: "Generation of Smurf2 Conditional Knockout Mice." in: International journal of biological sciences, Vol. 14, Issue 5, pp. 542-548, (2018) (PubMed).

Li, Chung, Li, Overstreet, Gagnon, Grouix, Leduc, Laurin, Zhang, Harris: "Fatty acid receptor modulator PBI-4050 inhibits kidney fibrosis and improves glycemic control." in: JCI insight, Vol. 3, Issue 10, (2018) (PubMed).

Li, Shi, Srivastava, Kitada, Nagai, Nitta, Kohno, Kanasaki, Koya: "FGFR1 is critical for the anti-endothelial mesenchymal transition effect of N-acetyl-seryl-aspartyl-lysyl-proline via induction of the MAP4K4 pathway." in: Cell death & disease, Vol. 8, Issue 8, pp. e2965, (2018) (PubMed).

Chung, Overstreet, Li, Wang, Niu, Wang, Fan, Sasaki, Jin, Khodo, Gewin, Zhang, Harris: "TGF-β promotes fibrosis after severe acute kidney injury by enhancing renal macrophage infiltration." in: JCI insight, Vol. 3, Issue 21, (2018) (PubMed).

Tang, Heller, Meng, Yu, Tang, Zhou, Zhang: "Transforming Growth Factor-β (TGF-β) Directly Activates the JAK1-STAT3 Axis to Induce Hepatic Fibrosis in Coordination with the SMAD Pathway." in: The Journal of biological chemistry, Vol. 292, Issue 10, pp. 4302-4312, (2017) (PubMed).

Subathra, Korrapati, Howell, Arthur, Shayman, Schnellmann, Siskind: "Kidney glycosphingolipids are elevated early in diabetic nephropathy and mediate hypertrophy of mesangial cells." in: American journal of physiology. Renal physiology, Vol. 309, Issue 3, pp. F204-15, (2015) (PubMed).

Luo, Xu, Chen, Warburton, Dong, Qian, Selman, Gauldie, Kolb, Shi: "A novel profibrotic mechanism mediated by TGFβ-stimulated collagen prolyl hydroxylase expression in fibrotic lung mesenchymal cells." in: The Journal of pathology, Vol. 236, Issue 3, pp. 384-94, (2015) (PubMed).

Herhaus, Al-Salihi, Macartney, Weidlich, Sapkota: "OTUB1 enhances TGFβ signalling by inhibiting the ubiquitylation and degradation of active SMAD2/3." in: Nature communications, Vol. 4, pp. 2519, (2014) (PubMed).

Kawamura, Maeda, Imamura, Setoguchi, Yokouchi, Ishidou, Komiya: "SnoN suppresses maturation of chondrocytes by mediating signal cross-talk between transforming growth factor-β and bone morphogenetic protein pathways." in: The Journal of biological chemistry, Vol. 287, Issue 34, pp. 29101-13, (2012) (PubMed).

Beckham, Tuttle, Tyler: "Reovirus activates transforming growth factor beta and bone morphogenetic protein signaling pathways in the central nervous system that contribute to neuronal survival following infection." in: Journal of virology, Vol. 83, Issue 10, pp. 5035-45, (2009) (PubMed).

Coffman, Coluccio, Planchart, Robertson: "Oral-aboral axis specification in the sea urchin embryo III. Role of mitochondrial redox signaling via H2O2." in: Developmental biology, Vol. 330, Issue 1, pp. 123-30, (2009) (PubMed).

Yamashita, Fatyol, Jin, Wang, Liu, Zhang: "TRAF6 mediates Smad-independent activation of JNK and p38 by TGF-beta." in: Molecular cell, Vol. 31, Issue 6, pp. 918-24, (2008) (PubMed).

Shi, Massagué: "Mechanisms of TGF-beta signaling from cell membrane to the nucleus." in: Cell, Vol. 113, Issue 6, pp. 685-700, (2003) (PubMed).

Target Details for SMAD3

Target: SMAD3 (SMAD, Mothers Against DPP Homolog 3 (SMAD3))

Alternative Name: SMAD3 (SMAD3 Products)

Background: Background: This antibody is designed, produced, and validated as part of a collaboration with 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.

Gene ID: 4088, 5174513

UniProt: P84022

Pathways: Cell Division Cycle, Chromatin Binding, Cell-Cell Junction Organization, Positive Regulation of Endopeptidase Activity, Autophagy