Anti-SMAD1+SMAD5 antibody [AF10B7] (ab75273)

Overview

  • Product nameAnti-SMAD1+SMAD5 antibody [AF10B7]
  • Description
    Mouse monoclonal [AF10B7] to SMAD1+SMAD5
  • Tested applicationsFlow Cyt, WB, ELISA more details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Recombinant human protein purified from E.coli (His-Smad1)

  • Positive control
    • SK-N-MC, Ramos, 293T and PC12 cell lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab75273 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
Flow Cyt Use 0.5µg for 106 cells.
WB 1/1000 - 1/2000. Predicted molecular weight: 52 kDa.
ELISA Use at an assay dependent dilution.

Target

  • RelevanceThe Smad family of proteins function in the transmission of extracellular signals in the TGF-ß signaling pathway. Binding of TGF-ß superfamily ligands to extracellular receptors triggers phosphorylation of Smad2 at a Serine-Serine-Methionine-Serine (SSMS) motif at its C-terminus. Phosphorylated Smad2 is then able to form a complex with Smad4. These complexes accumulate in the cell nucleus, where they directly participate in the regulation of gene expression. In mammals, eight Smad proteins have been identified to date. The Smad family of proteins can be divided into three functional groups: the receptor-activated Smads (R-Smads), common mediator Smads (Co-Smads), and the inhibitory Smads (I-Smads). The R-Smads are directly phosphorylated by the activated type I receptors on their C-terminal Ser-Ser-X-Ser (SSXS) motif and include Smad1, Smad2, Smad3, Smad5, and Smad8. Smad2 and Smad3 are phosphorylated in response to TGF-ß and activin, whereas Smad1, Smad5, and Smad8 are phosphorylated in response to BMP (Bone Morphogenetic Protein). This C-terminal phosphorylation allows R-Smad binding to Co-Smad, Smad4, and translocation to the nucleus where they regulate TGF-ß target genes. Smad6 and Smad7 belong to the I-Smad family which bind to the type I receptor or Smad4 and block their interaction with R-Smads.
  • Cellular localizationCytoplasm. Nucleus. Note=Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4.
  • Database links
  • Alternative names
    • MAD mothers against decapentaplegic homolog 1 antibody
    • MAD mothers against decapentaplegic homolog 5 antibody
    • MADH1 antibody
    • MADH5 antibody
    • Mothers against DPP homolog 1 antibody
    • Mothers against DPP homolog 5 antibody
    • SMAD family member 1 antibody
    • SMAD family member 5 antibody
    • SMAD1 antibody
    • SMAD5 antibody
    see all

Anti-SMAD1+SMAD5 antibody [AF10B7] images

  • All lanes : Anti-SMAD1+SMAD5 antibody [AF10B7] (ab75273) at 1/1000 dilution

    Lane 1 : SK-N-MC cell lysate
    Lane 2 : Ramos cell lysate
    Lane 3 : 293T cell lysate
    Lane 4 : PC12 cell lysate


    Predicted band size : 52 kDa
    Observed band size : 58 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 120 kDa. We are unsure as to the identity of these extra bands.
  • Overlay histogram showing HeLa cells stained with ab75273 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75273, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

References for Anti-SMAD1+SMAD5 antibody [AF10B7] (ab75273)

ab75273 has not yet been referenced specifically in any publications.

Product Wall

Thank you for contacting us.

Ab75273 can detect smad1 and smad5. It’s validated with each recombinant protein.

The molecular weight of smad1 and smad5 is same and that’s the reason why the predicted band size is same. Read More

Thank you for contacting us. Of the three antibodies we discussed yesterday, ab80255 is the best choice, given that we know the immunogen is derived from the N terminus, and your detection antibody is directed against the C terminus. The epitope that a...

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