Human/Mouse Smad2/3 Antibody

(7 citations)
(1 Review)
  
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human, mouse, and rat Smad2/3 in Western blots. Predicted to detect rat based on sequence homology.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Smad3
    Ser2-Ala230
    Accession # P84022
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    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
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.
Data Examples
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.
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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 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.

  • Long Name:
    Mothers Against DPP Homolog 2/3
  • Alternate Names:
    hMAD-2; hSMAD2; JV18; JV18-1SMAD, mothers against DPP homolog 2 (Drosophila); MAD homolog 2; Mad protein homolog; Mad, mothers against decapentaplegic homolog 2; MADH2MGC34440; MADR2MGC22139; Mad-related protein 2; mother against DPP homolog 2; mothers against decapentaplegic homolog 2; Mothers against DPP homolog 2; Sma- and Mad-related protein 2; SMAD 2; SMAD family member 2MAD, mothers against decapentaplegic homolog 2 (Drosophila); SMAD, mothers against DPP homolog 2; Smad2; Smad2/3
Related Research Areas
Citations:

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.

7 Citations: Showing 1 - 7
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Species
Applications
Sample Type
  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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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Average Rating: 5 (Based on 1 review)

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We have 1 review tested in 1 application: Western Blot.

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Images Ratings Applications Species Reviewed By Date Details
Western Blot Human/Mouse Smad2/3 Antibody AF3797
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 WB Human Anonymous 03/14/2017
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Western Blot Human/Mouse Smad2/3 Antibody AF3797
Western Blot: Human/Mouse Smad2/3 Antibody [AF3797]

Summary

ApplicationWestern Blot
Sample TestedHuman cell line
SpeciesHuman

Other Experimental Details

Other Experimental DetailsDetection 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.

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