Human/Mouse Smad2/3 Antibody Summary
Ser2-Ala230
Accession # P84022
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of Human/Mouse Smad2/3 by Western Blot. Western blot shows lysates of A549 human lung carcinoma cell line, Jurkat human acute T cell leukemia cell line, HEK293 human embryonic kidney cell line, and C2C12 mouse myoblast cell line. PVDF membrane was probed with 0.5 µg/mL Goat Anti-Human/Mouse Smad2/3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3797) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). For additional reference, recombinant human Smad2 and Smad3 were included. Specific bands for Smad2 were detected at approximately 64 and 58 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
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Detection of Smad2/3-regulated Genes by Chromatin Immunoprecipitation. Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin for 30 minutes was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Smad2/3/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human/Mouse Smad2/3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3797) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. Thec-mycpromoter was detected by standard PCR.
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Smad2/3 in MCF‑7 Human Cell Line. Smad2/3 was detected in immersion fixed MCF-7 human breast cancer cell line induced (upper panel) or non-induced (lower panel) to undergo epithelial-mesenchymal transition (EMT) using Goat Anti-Human/Mouse Smad2/3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3797) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and, in EMT-induced cells, nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
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Detection of Human Smad2/3 by Simple WesternTM. Simple Western lane view shows lysates of A549 human lung carcinoma cell line, COLO 205 human colorectal adenocarcinoma cell line, and HT-29 human colon adenocarcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Smad2/3 at approximately 64 kDa (as indicated) using 25 µg/mL of Goat Anti-Human/Mouse Smad2/3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3797) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
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Smad2/3 in Human Brain (Cortex). Smad2/3 was detected in immersion fixed paraffin-embedded sections of human brain (cortex) using Goat Anti-Human/Mouse Smad2/3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3797) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in neurons. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
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Smad2/3 in Human Brain (Cerebellum). Smad2/3 was detected in immersion fixed paraffin-embedded sections of human brain (cerebellum) using Goat Anti-Human/Mouse Smad2/3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3797) at 10 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in Purkinje neurons. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Smad2/3
Smads are a family of intracellular proteins that transmit transforming growth factor beta (TGF-beta ) superfamily signals from the cell surface to the nucleus. The Smad family is divided into three subclasses: receptor regulated Smads, (Smads 1, 2, 3, 5 and 8); the common partner, (Smad4); and the inhibitory Smads, (Smads 6 and 7). The binding of TGF-beta or activin to their cognate receptor induces phosphorylation of Smads 2 and 3. The activated Smads associate with the common-mediator subunit, Smad4, and the heteromeric complex translocates into the nucleus to initiate transcription. Smad3, also known as Mothers Against Decapentaplegic homolog 3 (MADH3), shares 83% amino acid identity with Smad2, also known as Mothers Against Decapentaplegic homolog 2 (MADH2). Human Smad2 has 99% identity to mouse and rat Smad2. Human Smad3 has 99% identity to mouse and rat Smad3.
Product Datasheets
Citations for Human/Mouse Smad2/3 Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
24
Citations: Showing 1 - 10
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KMT2A associates with PHF5A-PHF14-HMG20A-RAI1 subcomplex in pancreatic cancer stem cells and epigenetically regulates their characteristics
Authors: Mouti MA, Deng S, Pook M et al.
Nature communications
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TFEB-driven autophagy potentiates TGF-beta induced migration in pancreatic cancer cells
Authors: He R, Wang M, Zhao C et al.
J. Exp. Clin. Cancer Res.
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DPP-4 Inhibitors Attenuate Fibrosis After Glaucoma Filtering Surgery by Suppressing the TGF-?/Smad Signaling Pathway
Authors: Yoshida M, Kokubun T, Sato K et al.
Investigative ophthalmology & visual science
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The evolutionarily conserved long non‐coding RNA LINC00261 drives neuroendocrine prostate cancer proliferation and metastasis via distinct nuclear and cytoplasmic mechanisms
Authors: Rebecca L. Mather, Abhijit Parolia, Sandra E. Carson, Erik Venalainen, David Roig‐Carles, Mustapha Jaber et al.
Molecular Oncology
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Smooth Muscle Cell Reprogramming in Aortic Aneurysms
Authors: Chen PY, Qin L, Li G et al.
Cell Stem Cell
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RBL2-E2F-GCN5 guide cell fate decisions during tissue specification by regulating cell-cycle-dependent fluctuations of non-cell-autonomous signaling
Authors: Militi, S;Nibhani, R;Jalali, M;Pauklin, S;
Cell reports
Species: Human
Sample Types: Whole Cells
Applications: ICC -
Approaches in Hydroxytyrosol Supplementation on Epithelial-Mesenchymal Transition in TGFbeta1-Induced Human Respiratory Epithelial Cells
Authors: RA Razali, MD Yazid, A Saim, RBH Idrus, Y Lokanathan
International Journal of Molecular Sciences, 2023-02-16;24(4):.
Species: Human
Sample Types: Cell Lysates
Applications: Simple Western -
The Expression of Follistatin-like 1 Protein Is Associated with the Activation of the EMT Program in Sj�gren's Syndrome
Authors: M Sisto, D Ribatti, G Ingravallo, S Lisi
Journal of Clinical Medicine, 2022-09-13;11(18):.
Species: Human
Sample Types: Tissue Homogenates
Applications: Western Blot -
Immune modulation by complement receptor 3-dependent human monocyte TGF-beta1-transporting vesicles
Authors: LD Halder, EAH Jo, MZ Hasan, M Ferreira-G, T Krüger, M Westermann, DI Palme, G Rambach, N Beyersdorf, C Speth, ID Jacobsen, O Kniemeyer, B Jungnickel, PF Zipfel, C Skerka
Nat Commun, 2020-05-11;11(1):2331.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
TGFbeta1-Smad canonical and -Erk noncanonical pathways participate in interleukin-17-induced epithelial-mesenchymal transition in Sj�gren's syndrome
Authors: M Sisto, L Lorusso, G Ingravallo, D Ribatti, S Lisi
Lab. Invest., 2020-01-10;100(6):824-836.
Species: Human
Sample Types: Cell Lysates, Whole Cells
Applications: Flow Cytometry, Western Blot -
GDF3 Protects Mice against Sepsis-Induced Cardiac Dysfunction and Mortality by Suppression of Macrophage Pro-Inflammatory Phenotype
Authors: L Wang, Y Li, X Wang, P Wang, K Essandoh, S Cui, W Huang, X Mu, Z Liu, Y Wang, T Peng, GC Fan
Cells, 2020-01-03;9(1):.
Species: Mouse
Sample Types: Whole Cells
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Control over single-cell distribution of G1 lengths by WNT governs pluripotency
Authors: J Jang, D Han, M Golkaram, M Audouard, G Liu, D Bridges, S Hellander, A Chialastri, SS Dey, LR Petzold, KS Kosik
PLoS Biol., 2019-09-26;17(9):e3000453.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Maternal pluripotency factors initiate extensive chromatin remodelling to predefine first response to inductive signals
Authors: GE Gentsch, T Spruce, NDL Owens, JC Smith
Nat Commun, 2019-09-19;10(1):4269.
Species: Xenopus
Sample Types: Embryo, Tissue Homogenates
Applications: ChIP, IHC -
COA-Cl prevented TGF-?1-induced CTGF expression by Akt dephosphorylation in normal human dermal fibroblasts, and it attenuated skin fibrosis in mice models of systemic sclerosis
Authors: K Nakai, S Karita, J Igarashi, I Tsukamoto, K Hirano, Y Kubota
J. Dermatol. Sci., 2019-03-12;0(0):.
Species: Mouse
Sample Types: Cell Lysates
Applications: Western Blot -
The SMAD2/3 interactome reveals that TGF? controls m6A mRNA methylation in pluripotency
Authors: A Bertero, S Brown, P Madrigal, A Osnato, D Ortmann, L Yiangou, J Kadiwala, NC Hubner, IR de Los Moz, C Sadée, AS Lenaerts, S Nakanoh, R Grandy, E Farnell, J Ule, HG Stunnenber, S Mendjan, L Vallier
Nature, 2018-02-28;555(7695):256-259.
Species: Human
Sample Types: Protein
Applications: Immunoprecipitation -
Neuronal Protein 3.1 Deficiency Leads to Reduced Cutaneous Scar Collagen Deposition and Tensile Strength due to Impaired Transforming Growth Factor-?1 to -?3 Translation
Authors: Tao Cheng
Am. J. Pathol, 2016-12-08;0(0):.
Species: Mouse
Sample Types: Cell Lysates
Applications: Western Blot -
Potential mechanisms underlying ectodermal differentiation of Wharton's jelly mesenchymal stem cells
Biochem Biophys Res Commun, 2016-08-05;0(0):.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
HEB associates with PRC2 and SMAD2/3 to regulate developmental fates.
Authors: Yoon, Se-Jin, Foley, Joseph W, Baker, Julie C
Nat Commun, 2015-03-16;6(0):6546.
Species: Mouse
Sample Types: Whole Cells
Applications: ChIP -
miR-373 is regulated by TGFbeta signaling and promotes mesendoderm differentiation in human Embryonic Stem Cells.
Authors: Rosa A, Papaioannou M, Krzyspiak J, Brivanlou A
Dev Biol, 2014-04-04;391(1):81-8.
Species: Human
Sample Types: Whole Cells
Applications: ChIP -
Antagonism of Nodal signaling by BMP/Smad5 prevents ectopic primitive streak formation in the mouse amnion.
Authors: Pereira P, Dobreva M, Maas E, Cornelis F, Moya I, Umans L, Verfaillie C, Camus A, de Sousa Lopes S, Huylebroeck D, Zwijsen A
Development, 2012-09-01;139(18):3343-54.
Species: Human
Sample Types: Cell Lysates
Applications: Immunoprecipitation -
Satellite cell senescence underlies myopathy in a mouse model of limb-girdle muscular dystrophy 2H.
Authors: Kudryashova E, Kramerova I, Spencer MJ
J. Clin. Invest., 2012-04-16;122(5):1764-76.
Species: Mouse
Sample Types: Tissue Homogenates
Applications: Western Blot -
Chromatin and transcriptional signatures for Nodal signaling during endoderm formation in hESCs.
Authors: Kim SW, Yoon SJ, Chuong E, Oyolu C, Wills AE, Gupta R, Baker J
Dev. Biol., 2011-06-29;357(2):492-504.
Species: Human
Sample Types: Cell Lysates
Applications: ChIP -
The transforming growth factor-beta/Smad2,3 signalling axis is impaired in experimental pulmonary hypertension.
Authors: Zakrzewicz A, Kouri FM, Nejman B, Kwapiszewska G, Hecker M, Sandu R, Dony E, Seeger W, Schermuly RT, Eickelberg O, Morty RE
Eur. Respir. J., 2007-03-28;29(6):1094-104.
Species: Rat
Sample Types: Whole Tissue
Applications: IHC -
Comprehensive characterization of chorionic villi-derived mesenchymal stromal cells from human placenta
Authors: MS Ventura Fe, M Bienert, K Müller, B Rath, T Goecke, C Opländer, T Braunschwe, P Mela, TH Brümmendor, F Beier, S Neuss
Stem Cell Res Ther, 2018-02-05;9(1):28.
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Detection of Human Smad 2/3 in melanoma and mammary tumor cell lines. Dilution: 1:500 in PBS with 5% BSA. Secondary Ab: anti-Goat IgG 1:5,000.
