Overview

  • Product nameAnti-Rad53 antibody
    See all Rad53 primary antibodies
  • Description
    Rabbit polyclonal to Rad53
  • Tested applicationsSuitable for: WBmore details
  • Species reactivity
    Reacts with: Saccharomyces cerevisiae
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 700 to the C-terminus of Saccharomyces cerevisiae Rad53.

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: TCA preps from exponentially growing WT yeast cells.; TCA preps from exponentially growing WT yeast cells treated with Hydroxyurea (HU).; TCA preps from exponentially growing rad53delta yeast cells.

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 bufferpH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS
    Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab104232 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 1/2000. Detects a band of approximately 92 kDa (predicted molecular weight: 92 kDa). Abcam recommends using milk as the blocking agent (4%)

Target

  • FunctionControls S-phase checkpoint as well as G1 and G2 DNA damage checkpoints. Phosphorylates proteins on serine, threonine, and tyrosine. Prevents entry into anaphase and mitotic exit after DNA damage via regulation of the Polo kinase CDC5. Seems to be involved in the phosphorylation of RPH1.
  • Sequence similaritiesBelongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CHEK2 subfamily.
    Contains 2 FHA domains.
    Contains 1 protein kinase domain.
  • DomainFHA domains are phosphothreonine recognition modules, FHA 1 strongly selects for Asp at position +3 relative to phosphothreonine, whereas FHA 2 selects for Ile in this position.
  • Post-translational
    modifications
    Autophosphorylated.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Database links
    • Alternative names
      • CHEK2 homolog antibody
      • CHK2 homolog antibody
      • MEC2 antibody
      • RAD53 antibody
      • RAD53_YEAST antibody
      • Serine protein kinase 1 antibody
      • Serine-protein kinase 1 antibody
      • Serine/threonine-protein kinase RAD53 antibody
      • SPK1 antibody
      see all

    Anti-Rad53 antibody images

    • All lanes : Anti-Rad53 antibody (ab104232) at 1/2000 dilution

      Lane 1 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells- TCA prep.
      Lane 2 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells treated with 20mg/ml phleomycin for 1 hour - TCA prep
      Lane 3 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells- TCA prep
      Lane 4 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells arrested in G2 phase with nocodazole- TCA prep.
      Lane 5 : Saccharomyces cerevisiae whole cell lysate from exponentially growing cells arrested in G2 phase with nocodazole and treated with 20mg/ml phleomycin- TCA prep.

      Lysates/proteins at 20 µg per lane.

      Secondary
      Amersham anti-rabbit IgG conjugated to HRP at 1/10000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 92 kDa
      Observed band size : 92 kDa
      Additional bands at : >92 kDa (possible post-translational modification).

      Exposure time : 2 minutes

      Michela Clerici, University of Milano-Biococca

      Blocking: 4% milk for 1 hour

      Phosphorylation of Rad53 induced by phleomycin treatment see PubmedID:152856

      See Abreview

    • All lanes : Anti-Rad53 antibody (ab104232) at 1/2000 dilution

      Lane 1 : TCA preps from exponentially growing WT yeast cells.
      Lane 2 : TCA preps from exponentially growing WT yeast cells treated with Hydroxyurea (HU).
      Lane 3 : TCA preps from exponentially growing rad53delta yeast cells.

      Secondary
      1/10000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 92 kDa


      Exposure time : 30 seconds

      This image is courtesy of Ilaria Guerini, Gurdon Institute, United Kingdom

    • All lanes : Anti-Rad53 antibody (ab104232) at 1/1000 dilution

      Lane 1 : Alpha-factor arrested WT cells.
      Lane 2 : WT cells 20' after release in S-phase.
      Lane 3 : WT cells 40' after release in S-phase.
      Lane 4 : WT cells 60' after release in S-phase.
      Lane 5 : Alpha-factor arrested WT cells.
      Lane 6 : WT cells 20' after release in S-phase in 200mM HU.
      Lane 7 : WT cells 40' after release in S-phase in 200mM HU.
      Lane 8 : WT cells 60' after release in S-phase in 200mM HU.
      Lane 9 : Alpha-factor arrested rad53delta cells.
      Lane 10 : rad53delta cells 20' after release in S-phase.
      Lane 11 : rad53delta cells 40' after release in S-phase.
      Lane 12 : rad53delta cells 60' after release in S-phase.
      Lane 13 : Alpha-factor arrested rad53delta cells.
      Lane 14 : rad53delta cells 20' after release in S-phase in 200mM HU.
      Lane 15 : rad53delta cells 40' after release in S-phase in 200mM HU.
      Lane 16 : rad53delta cells 60' after release in S-phase in 200mM HU.

      Secondary
      Donkey Anti-Rabbit IgG H&L (HRP) (ab16284) at 1/2000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 92 kDa
      Additional bands at : above 92 kDa (possible post-translational modification).

      Exposure time : 2 minutes

      See Abreview


    • Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 92 kDa


      Exposure time : 1 minute
      Western blot showing ab104232 detecting Rad53 in both its unphosphorylated and phosphorylated status. ab104232 was used at 1:2000 dilution.

      See Abreview

    References for Anti-Rad53 antibody (ab104232)

    This product has been referenced in:
    • Wang SH  et al. Curcumin-Mediated HDAC Inhibition Suppresses the DNA Damage Response and Contributes to Increased DNA Damage Sensitivity. PLoS One 10:e0134110 (2015). Read more (PubMed: 26218133) »
    • Jha DK & Strahl BD An RNA polymerase II-coupled function for histone H3K36 methylation in checkpoint activation and DSB repair. Nat Commun 5:3965 (2014). Read more (PubMed: 24910128) »

    See all 4 Publications for this product

    Product Wall

    Western blot

    Excellent
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing (10%)
    Sample Saccharomyces cerevisiae Cell lysate - whole cell (S. cerevisiae cells)
    Specification S. cerevisiae cells
    Treatment Exponentially growing or G2-arrested cells treated with Phleomycin
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
    Username

    Dr. Michela Clerici

    Verified customer

    Submitted May 07 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Saccharomyces cerevisiae Cell lysate - whole cell (whole cell extract (done by TCA 10% protocol))
    Loading amount 15 µg
    Specification whole cell extract (done by TCA 10% protocol)
    Treatment cells were grown in galactose 2% to induce HO nuclease
    Gel Running Conditions Reduced Denaturing (gel 10%)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Dr. Achille Pellicioli

    Verified customer

    Submitted Nov 09 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Saccharomyces cerevisiae Cell lysate - whole cell (W303)
    Loading amount 2e+006 cells
    Specification W303
    Treatment release into 200mM HU
    Gel Running Conditions Reduced Denaturing (7.5%)
    Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Jul 31 2012

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"