Product nameAnti-SMAD1+SMAD5 antibody [AF10B7]
DescriptionMouse monoclonal [AF10B7] to SMAD1+SMAD5
Tested applicationsSuitable for: ChIP, Flow Cyt, WB, ELISAmore details
Species reactivityReacts with: Rat, Human
Recombinant human protein purified from E.coli (His-Smad1)
- SK-N-MC, Ramos, 293T and PC12 cell lysates.
This product was changed from ascites to tissue culture supernatant on 18th September 2017. Lot numbers higher than GR188031 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.
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, 0.87% Sodium chloride, HEPES, 0.01% BSA
PurityTissue culture supernatant
Light chain typekappa
Our Abpromise guarantee covers the use of ab75273 in the following tested applications.
|ChIP||Use at an assay dependent concentration.|
|Flow Cyt||Use at an assay dependent concentration.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 52 kDa.|
|ELISA||Use at an assay dependent concentration.|
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.
- MAD mothers against decapentaplegic homolog 1 antibody
- MAD mothers against decapentaplegic homolog 5 antibody
- MADH1 antibody
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.
This product has been referenced in:
- Wen Q et al. 8-bromo-7-methoxychrysin suppress stemness of SMMC-7721 cells induced by co-culture of liver cancer stem-like cells with hepatic stellate cells. BMC Cancer 19:224 (2019). Read more (PubMed: 30866863) »
- Zhang Y et al. Selective compounds enhance osteoblastic activity by targeting HECT domain of ubiquitin ligase Smurf1. Oncotarget 8:50521-50533 (2017). Read more (PubMed: 28881580) »