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 controlSK-N-MC, Ramos, 293T and PC12 cell lysates.

Properties

• FormLiquid
• Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
• Storage bufferPreservative: 0.03% Sodium Azide
  Constituents: 50% Glycerol, 0.01% BSA, HEPES, 0.15M Sodium chloride
• Concentration information loading...
• PurityAmmonium Sulphate Precipitation
• Clonality Monoclonal
• Clone numberAF10B7
• IsotypeIgG2a
• Light chain typekappa
• Research Areas
  • Stem Cells
  • Signaling Pathways
  • TGF beta
  • Cytoplasmic

  • Cancer
  • Signal transduction
  • Nuclear signaling
  • SMAD family

  • Signal Transduction
  • Signaling Pathway
  • Nuclear Signaling
  • SMADs

  • Epigenetics and Nuclear Signaling
  • Nuclear Signaling Pathways
  • SMADs

Applications

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
  • Entrez Gene: 4086 Human
  • Entrez Gene: 4090 Human
  • Entrez Gene: 25671 Rat
  • Entrez Gene: 59328 Rat
  • Omim: 601595 Human
  • Omim: 603110 Human
  • SwissProt: Q15797 Human
  • SwissProt: Q99717 Human

  • SwissProt: P97588 Rat
  • SwissProt: Q9R1V3 Rat

  see all

• Alternative names
  MAD mothers against decapentaplegic homolog 1 antibodyMAD mothers against decapentaplegic homolog 5 antibodyMADH1 antibody

  MADH5 antibodyMothers against DPP homolog 1 antibodyMothers against DPP homolog 5 antibodySMAD family member 1 antibodySMAD family member 5 antibodySMAD1 antibodySMAD5 antibody

  see all

Anti-SMAD1+SMAD5 antibody [AF10B7] images

• 

  Western blot - SMAD1+SMAD5 antibody [AF10B7] (ab75273)
  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.
• 

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