Human Phospho-Smad2/3 (S465/S467) Antibody

Recombinant Monoclonal Antibody.
  
  • Species Reactivity
    Human
  • Specificity
    Detects human Smad2/3 when dually phosphorylated at S465/S467 in Western blots.
  • Source
    Recombinant Monoclonal Rabbit IgG Clone # 1074A
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    Phosphopeptide containing the human Smad2 S465/S467 site
    Accession # NP_005892
  • 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
    2 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 Phospho-Smad2/3 (S465/S467) by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with 10 ng/mL Recombinant Human TGF‑ beta 1 (Catalog # 240-B) for 30 minutes. PVDF membrane was probed with 2 μg/mL of Rabbit Anti-Human Phospho-Smad2/3 (S465/S467) Monoclonal Antibody (Catalog # MAB8935) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-Smad2/3 (S465/S467) at approximately 68 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
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

Smad2 (Sma and MAD-related Protein 2), also known as MAD2, MADR2 and MADH2, is a 58-62 kDa member of the Dwarfin/SMAD family of proteins. It is widely expressed, particularly in striated muscle, and exists constitutively in the cytoplasm. Smad2 is a downstream mediator of TGF-beta and activin signaling. In particular, Smad2 is a nonphosphorylated monomer in unstimulated cells. Upon ALK-4, -5, and -7 receptor activation, Smad2 is phosphorylated and forms either homotrimers or heterotrimers with Smad3 and Smad4. These heterotrimers enter the nucleus and initiate gene transcription. Human Smad2 is 467 amino acids (aa) in length. It contains one regulatory MH domain (aa 120-176) and a transactivation MH domain (aa 274-467). There are at least five utilized phosphorylation sites and one acetylation site at Lys19 that promotes transcriptional activity. There are four potential isoform variants. One shows a deletion of aa 79-108, while another contains the same deletion coupled to another deletion of aa 219-243. A third shows a deletion of aa 344-358, and a fourth is missing aa 221-225. Over aa 20-108, human Smad2 shares 98% aa identity with mouse Smad2.

  • 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
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Secondary Antibodies
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Rabbit IgG HRP-conjugated Antibody

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Rabbit IgG APC-conjugated Antibody

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Rabbit IgG PE-conjugated Antibody

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Rabbit IgG Fluorescein-conjugated Antibody

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Donkey Anti-Rabbit IgG NorthernLights™ NL493-conjugated Antibody

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Goat Anti-Rabbit IgG Biotinylated Antibody

WB BAF008 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Rabbit IgG Antibody

WB AF008 1
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Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC003  
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Donkey Anti-Rabbit IgG (H+L) Antibody

D-301-C-ABS2
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