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  1. Link

    pe-smad3-antibody-ep568y-ab208751.pdf

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Signal Transduction Signaling Pathway Nuclear Signaling SMADs
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RecombinantRabMAb

Recombinant PE Anti-Smad3 antibody [EP568Y] (ab208751)

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Flow Cytometry - PE Anti-Smad3 antibody [EP568Y] (ab208751)
  • Immunocytochemistry/ Immunofluorescence - PE Anti-Smad3 antibody [EP568Y] (ab208751)
  • PE Anti-Smad3 antibody [EP568Y] (ab208751)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • PE Rabbit monoclonal [EP568Y] to Smad3
  • Suitable for: ICC/IF, Flow Cyt
  • Reacts with: Human
  • Conjugation: PE. Ex: 488nm, Em: 575nm

You may also be interested in

Protein
Product image
Recombinant Human Smad3 protein (ab89353)

View more associated products

Overview

  • Product name

    PE Anti-Smad3 antibody [EP568Y]
    See all Smad3 primary antibodies
  • Description

    PE Rabbit monoclonal [EP568Y] to Smad3
  • Host species

    Rabbit
  • Conjugation

    PE. Ex: 488nm, Em: 575nm
  • Tested applications

    Suitable for: ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide within Human Smad3 aa 200-300 (internal sequence). The exact sequence is proprietary.
    Database link: P84022
    (Peptide available as ab173094)

  • Positive control

    • Flow Cyt: HepG2 cells ICC/IF: HepG2 cells
  • General notes

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot. Store at +4°C. Do Not Freeze. Store In the Dark.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 1% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP568Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • SMADs
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • SMADs
    • Stem Cells
    • Signaling Pathways
    • TGF beta
    • Cytoplasmic
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • SMAD family
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia

Associated products

  • Alternative Versions

    • Anti-Smad3 antibody [EP568Y] - BSA and Azide free (ab157372)
    • Alexa Fluor® 488 Anti-Smad3 antibody [EP568Y] (ab204257)
    • Alexa Fluor® 647 Anti-Smad3 antibody [EP568Y] (ab204461)
    • HRP Anti-Smad3 antibody [EP568Y] (ab204462)
    • Alexa Fluor® 594 Anti-Smad3 antibody [EP568Y] (ab206350)
    • Alexa Fluor® 555 Anti-Smad3 antibody [EP568Y] (ab206582)
    • Anti-Smad3 antibody [EP568Y] (ab40854)
  • Isotype control

    • PE Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab209478)
  • Recombinant Protein

    • Recombinant Human Smad3 protein (ab89353)

Applications

Our Abpromise guarantee covers the use of ab208751 in the following tested applications.

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

Application Abreviews Notes
ICC/IF 1/1000.

This product gave a positive signal in HepG2 cells fixed with 4% formaldehyde (10 min)

Flow Cyt 1/500.

Target

  • Function

    Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD3/SMAD4 complex, activates transcription. Also can form a SMAD3/SMAD4/JUN/FOS complex at the AP-1/SMAD site to regulate TGF-beta-mediated transcription. Has an inhibitory effect on wound healing probably by modulating both growth and migration of primary keratinocytes and by altering the TGF-mediated chemotaxis of monocytes. This effect on wound healing appears to be hormone-sensitive. Regulator of chondrogenesis and osteogenesis and inhibits early healing of bone fractures. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.
  • Involvement in disease

    Colorectal cancer
    Loeys-Dietz syndrome 3
  • Sequence similarities

    Belongs to the dwarfin/SMAD family.
    Contains 1 MH1 (MAD homology 1) domain.
    Contains 1 MH2 (MAD homology 2) domain.
  • Domain

    The MH1 domain is required for DNA binding. Also binds zinc ions which are necessary for the DNA binding.
    The MH2 domain is required for both homomeric and heteromeric interactions and for transcriptional regulation. Sufficient for nuclear import.
    The linker region is required for the TGFbeta-mediated transcriptional activity and acts synergistically with the MH2 domain.
  • Post-translational
    modifications

    Phosphorylated on serine and threonine residues. Enhanced phosphorylation in the linker region on Thr-179, Ser-204 and Ser-208 on EGF and TGF-beta treatment. Ser-208 is the main site of MAPK-mediated phosphorylation. CDK-mediated phosphorylation occurs in a cell-cycle dependent manner and inhibits both the transcriptional activity and antiproliferative functions of SMAD3. This phosphorylation is inhibited by flavopiridol. Maximum phosphorylation at the G(1)/S junction. Also phosphorylated on serine residues in the C-terminal SXS motif by TGFBR1 and ACVR1. TGFBR1-mediated phosphorylation at these C-terminal sites is required for interaction with SMAD4, nuclear location and transactivational activity, and appears to be a prerequisite for the TGF-beta mediated phosphorylation in the linker region. Dephosphorylated in the C-terminal SXS motif by PPM1A. This dephosphorylation disrupts the interaction with SMAD4, promotes nuclear export and terminates TGF-beta-mediated signaling. Phosphorylation at Ser-418 by CSNK1G2/CK1 promotes ligand-dependent ubiquitination and subsequent proteasome degradation, thus inhibiting SMAD3-mediated TGF-beta responses. Phosphorylated by PDPK1.
    Acetylation in the nucleus by EP300 in the MH2 domain regulates positively its transcriptional activity and is enhanced by TGF-beta.
    Ubiquitinated. Monoubiquitinated, leading to prevent DNA-binding. Deubiquitination by USP15 alleviates inhibition and promotes activation of TGF-beta target genes.
    Poly-ADP-ribosylated by PARP1 and PARP2. ADP-ribosylation negatively regulates SMAD3 transcriptional responses during the course of TGF-beta signaling.
  • Cellular localization

    Cytoplasm. Nucleus. Cytoplasmic and nuclear in the absence of TGF-beta. On TGF-beta stimulation, migrates to the nucleus when complexed with SMAD4 (PubMed:15799969). Through the action of the phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1 (PubMed:16751101, PubMed:19289081). Co-localizes with LEMD3 at the nucleus inner membrane (PubMed:15601644). MAPK-mediated phosphorylation appears to have no effect on nuclear import (PubMed:19218245). PDPK1 prevents its nuclear translocation in response to TGF-beta (PubMed:17327236).
  • Target information above from: UniProt accession P84022 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4088 Human
    • Entrez Gene: 17127 Mouse
    • Entrez Gene: 25631 Rat
    • Omim: 603109 Human
    • SwissProt: P84022 Human
    • SwissProt: Q8BUN5 Mouse
    • SwissProt: P84025 Rat
    • Unigene: 727986 Human
    • Unigene: 742270 Human
    • Unigene: 7320 Mouse
    • Unigene: 10636 Rat
    see all
  • Alternative names

    • DKFZP586N0721 antibody
    • DKFZp686J10186 antibody
    • hMAD 3 antibody
    • hMAD-3 antibody
    • hSMAD3 antibody
    • HSPC193 antibody
    • HST17436 antibody
    • JV15 2 antibody
    • JV15-2 antibody
    • JV152 antibody
    • LDS1C antibody
    • LDS3 antibody
    • MAD (mothers against decapentaplegic Drosophila) homolog 3 antibody
    • MAD homolog 3 antibody
    • Mad homolog JV15 2 antibody
    • Mad protein homolog antibody
    • MAD, mothers against decapentaplegic homolog 3 antibody
    • Mad3 antibody
    • MADH 3 antibody
    • MADH3 antibody
    • MGC60396 antibody
    • Mothers against decapentaplegic homolog 3 antibody
    • Mothers against DPP homolog 3 antibody
    • SMA and MAD related protein 3 antibody
    • SMAD 3 antibody
    • SMAD antibody
    • SMAD family member 3 antibody
    • SMAD, mothers against DPP homolog 3 antibody
    • Smad3 antibody
    • SMAD3_HUMAN antibody
    see all

Images

  • Flow Cytometry - PE Anti-Smad3 antibody [EP568Y] (ab208751)
    Flow Cytometry - PE Anti-Smad3 antibody [EP568Y] (ab208751)

    Overlay histogram showing HepG2 cells stained with ab208751 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 90% methanol (-20°C) for 30 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab208751, 1/500 dilution) for 30 min at 22°C.

    Isotype control antibody (black line) was rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20 mW Solid State Blue Laser (488nm) and 585/40 bandpass filter

  • Immunocytochemistry/ Immunofluorescence - PE Anti-Smad3 antibody [EP568Y] (ab208751)
    Immunocytochemistry/ Immunofluorescence - PE Anti-Smad3 antibody [EP568Y] (ab208751)

    ab208751 staining Smad3 in HepG2 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab208751 at 1/1000 dilution (pseudocolored in green) and ab195884, Rat monoclonal to Tubulin (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • PE Anti-Smad3 antibody [EP568Y] (ab208751)
    PE Anti-Smad3 antibody [EP568Y] (ab208751)

Protocols

  • Flow cytometry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (0)

    Publishing research using ab208751? Please let us know so that we can cite the reference in this datasheet.

    ab208751 has not yet been referenced specifically in any publications.

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