Overview

  • Product nameAnti-Scc4 antibody
    See all Scc4 primary antibodies
  • Description
    Rabbit polyclonal to Scc4
  • Tested applicationsSuitable for: WB, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Xenopus laevis
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 550 to the C-terminus of Human Scc4.

    (Peptide available as ab61625.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysate: HeLa

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferPreservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab46906 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 69 kDa (predicted molecular weight: 69 kDa).
IP Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • RelevanceScc4 (Mau2) is the metazoan homologue of Scc4 in S. cerevisiae. In yeast, Scc4 binds to Scc2 to form an essential complex that loads cohesion onto chromosomes. Similarly, the Scc4 / Mau2 protein in metazoans plays a significant role in sister chromatid separation and in loading of cohesin onto chromatin. The Scc2-Scc4 complex is tethered to pre-replication complexes through the Cdc7-Drf1 protein kinase DDK.
  • Cellular localizationNuclear
  • Database links
  • Alternative names
    • K0892 antibody
    • KIAA0892 antibody
    • Mau 2 antibody
    • Mau 2L antibody
    • mau-2 antibody
    • Mau2 antibody
    • MAU2L antibody
    • Scc 4 antibody
    • SCC4 antibody
    • sister chromatid cohesion 4 antibody
    see all

Anti-Scc4 antibody images

  • ICC/IF image of ab46906 stained HeLa cells. The cells were 100% methanol fixed (5 min) then permeabilised using 0.1% PBS-Triton and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab46906 at 1µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • All lanes : Anti-Scc4 antibody (ab46906) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate

    Lysates/proteins at 30 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 69 kDa
    Observed band size : 69 kDa
    Additional bands at : 48 kDa,80-100 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 10 minutes
  • Scc4 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Scc4 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab46906.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 69kDa: Scc4; non specific - 53kDa: We are unsure as to the identity of this extra band.
  • Lanes 1 - 2 : Blank
    Lanes 3 - 4 : Anti-Scc4 antibody (ab46906) at 1 µg/ml
    Lane 5 : Custom-made antibody at 1 µg/ml
    Lane 6 : Custom-made antibody at 1 µg/ml
    Lanes 7 - 8 : Secondary antibody only

    Lane 1 : HeLa cell lysate (40 µg)
    Lane 2 : HeLa cell lysate (20 µg)
    Lane 3 : HeLa cell lysate (40 µg)
    Lane 4 : HeLa cell lysate (20 µg)
    Lane 5 : HeLa cell lysate (40 µg)
    Lane 6 : HeLa cell lysate (20 µg)
    Lane 7 : HeLa cell lysate (40 µg)
    Lane 8 : HeLa cell lysate (20 µg)

    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 69 kDa
    Observed band size : 69 kDa


    Exposure time : 15 seconds

    Image kindly provided by Vlad Seitan, Institute of Human Genetics, Newcastle, United Kingdom

    Western blot showing detection of Scc4 in HeLa cell lysate by ab46906. Bands marked * correspond to the expected band for Scc4. The custom-made antibody was characterised by Vlad Seitan and co-workers.

References for Anti-Scc4 antibody (ab46906)

ab46906 has not yet been referenced specifically in any publications.

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