Human/Mouse Smad2/3 Antibody

Catalog # Availability Size / Price Qty
AF3797
AF3797-SP
Detection of Human/Mouse Smad2/3 by Western Blot.
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Product Details
Citations (10)
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Supplemental Products
Reviews (1)

Human/Mouse Smad2/3 Antibody Summary

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

Western Blot Detection of Human/Mouse Smad2/3 by Western Blot. View Larger

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. View Larger

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. View Larger

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. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

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.

10 Citations: Showing 1 - 10
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  1. 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;0(0):.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  2. 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 Types: Protein
    Applications: Immunoprecipitation
  3. Potential mechanisms underlying ectodermal differentiation of Wharton's jelly mesenchymal stem cells
    Biochem Biophys Res Commun, 2016;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  4. 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 Types: Cell Lysates
    Applications: Western Blot
  5. 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 Types: Whole Cells
    Applications: ChIP
  6. 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 Types: Whole Cells
    Applications: ChIP
  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 Types: Tissue Homogenates
    Applications: Western Blot
  8. 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 Types: Cell Lysates
    Applications: Immunoprecipitation
  9. 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 Types: Cell Lysates
    Applications: ChIP
  10. 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 Types: Whole Tissue
    Applications: IHC

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Reviews for Human/Mouse Smad2/3 Antibody

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Human/Mouse Smad2/3 Antibody
By Anonymous on 03/14/2017
Application: WB Sample Tested: Human cell line Species: Human

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.