抗Smad2/3抗体(Anti-Smad2/3, Goat-Poly antibody)

• 価格
• Product Details
• Applications and Data
• Related Product & Information
• Citations

価格

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Product Details

Species Reactivity	Human, Mouse
Label	Unconjugated
Immunogen	E. coli-derived recombinant human Smad3Ser2-Ala230Accession # P84022
Source	Polyclonal Goat IgG
Purification	Antigen Affinity-purified
Specificity	Detects human, mouse, and rat Smad2/3 in Western blots. Predicted to detect rat based on sequence homology.

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Applications and Data

	Recommended Concentration	Sample
Western Blot	0.5 µg/mL	See below
Simple Western	25 µg/mL	See below
Chromatin Immunoprecipitation (ChIP)	5 µg/5 x 106 cells	See below
Immunocytochemistry	5-15 µg/mL	See below

Western Blot
	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.
Chromatin Immunoprecipitation (ChIP)
	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. The c-myc promoter was detected by standard PCR.
Immunocytochemistry
	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.
Simple Western
	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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Related Product & Information

Long Name	Mothers Against DPP Homolog 2/3
Background	Smad2/3
background_content	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.

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Citations

1. 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;555(7695):256-259.
   Species: Human
   Sample Type: Protein
   Application: IP


2. Potential mechanisms underlying ectodermal differentiation of Wharton's jelly mesenchymal stem cells
   Biochem Biophys Res Commun, 2016;0(0):.
   Species: Human
   Sample Type: Cell Lysates
   Application: WB


3. 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;0(0):.
   Species: Mouse
   Sample Type: Cell Lysates
   Application: WB


4. HEB associates with PRC2 and SMAD2/3 to regulate developmental fates.
   Authors: Yoon, Se-Jin, Foley, Joseph W, Baker, Julie C
   Nat Commun, 2015;6(0):6546.
   Species: Mouse
   Sample Type: Whole Cells
   Application: ChIP


5. 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;391(1):81-8.
   Species: Human
   Sample Type: Whole Cells
   Application: ChIP


6. 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;139(18):3343-54.
   Species: Human
   Sample Type: Cell Lysates
   Application: IP


7. 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;122(5):1764-76.
   Species: Mouse
   Sample Type: Tissue Homogenates
   Application: WB


8. 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;357(2):492-504.
   Species: Human
   Sample Type: Cell Lysates
   Application: ChIP


9. 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;29(6):1094-104.
   Species: Rat
   Sample Type: Whole Tissue
   Application: IHC

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