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Human SMURF2 knockout HeLa cell pellet (ab279225)

Overview

  • Product name

    Human SMURF2 knockout HeLa cell pellet
    See all SMURF 2 kits

  • Product overview

    Abcam’s knockout cell pellets give you access to native proteins, without the need to culture cells. Our knockout cell pellets are prepared from our single-gene knockout cell lines and provide an additional offering to our cell lysates.

    Cells are snap-frozen to provide high quality pellets that are suitable for extraction with alternative lysis buffers or for preparation of lysates from subcellular fractions. Our knockout cell pellets are suitable for a variety of applications, including PCR, gene expression profiling and DNA library preparation.

  • Parental Cell Line

    HeLa

  • Organism

    Human

  • Mutation description

    Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon5 and 2 bp deletion in exon5.

  • Passage number

  • Knockout validation

    Sanger Sequencing, Western Blot (WB)

  • Notes

    Pellet size: 5 million cells/vial.

    This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

  • Tested applications

    Suitable for: WBmore details

Properties

Target

  • Function

    E3 ubiquitin-protein ligase which accepts ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then directly transfers the ubiquitin to targeted substrates. Interacts with SMAD1 and SMAD7 in order to trigger their ubiquitination and proteasome-dependent degradation. In addition, interaction with SMAD7 activates autocatalytic degradation, which is prevented by interaction with SCYE1. Forms a stable complex with the TGF-beta receptor-mediated phosphorylated SMAD2 and SMAD3. In this way, SMAD2 may recruit substrates, such as SNON, for ubiquitin-mediated degradation. Enhances the inhibitory activity of SMAD7 and reduces the transcriptional activity of SMAD2. Coexpression of SMURF2 with SMAD1 results in considerable decrease in steady-state level of SMAD1 protein and a smaller decrease of SMAD2 level.

  • Tissue specificity

    Widely expressed.

  • Pathway

    Protein modification; protein ubiquitination.

  • Sequence similarities

    Contains 1 C2 domain.
    Contains 1 HECT (E6AP-type E3 ubiquitin-protein ligase) domain.
    Contains 3 WW domains.

  • Domain

    The second and third WW domains are responsible for interaction with the PY-motif of R-SMAD (SMAD1, SMAD2 and SMAD3).
    The C2 domain is involved in autoinhibition of the catalytic activity by interacting with the HECT domain.

  • Post-translational
    modifications

    Auto-ubiquitinated and ubiquitinated in the presence of RNF11 and UBE2D1.

  • Cellular localization

    Nucleus. Cytoplasm. Cell membrane. Membrane raft. Cytoplasmic in the presence of SMAD7. Co-localizes with CAV1, SMAD7 and TGF-beta receptor in membrane rafts.
  • Information by UniProt

  • Alternative names

    • E3 ubiquitin-protein ligase SMURF2
    • EC 6.3.2.
    • hSMURF2
    • MGC138150
    • Smad specific E3 ubiquitin ligase 2
    • SMAD specific E3 ubiquitin protein ligase 2
    • SMAD ubiquitination regulatory factor 2
    • SMAD-specific E3 ubiquitin-protein ligase 2
    • SMUF2_HUMAN
    • Smurf2
    • Ubiquitin protein ligase SMURF2
    see all

Properties

Images

  • Western blot - Human SMURF2 knockout HeLa cell pellet (ab279225)
    Western blot - Human SMURF2 knockout HeLa cell pellet (ab279225)

    Lane 1:Wild-type HeLa cell lysate (20 ug)
    Lane 2:SMURF2 knockout HeLa cell lysate (20 ug)
    Lane 3:SH-SY5Y cell lysate (20 ug)

    ab53316 was shown to specifically react with SMURF 2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265231 (knockout cell lysate ab257691) was used. Wild-type and SMURF 2 knockout samples were subjected to SDS-PAGE. ab53316 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Sanger Sequencing - Human SMURF2 knockout HeLa cell pellet (ab279225)
    Sanger Sequencing - Human SMURF2 knockout HeLa cell pellet (ab279225)

    Allele-1: 2 bp deletion in exon5

     

  • Sanger Sequencing - Human SMURF2 knockout HeLa cell pellet (ab279225)
    Sanger Sequencing - Human SMURF2 knockout HeLa cell pellet (ab279225)

    Allele-2: 1 bp insertion in exon5

     

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