Overview

  • Product name

    Anti-Chk2 (phospho T68) antibody [Y171]
    See all Chk2 primary antibodies
  • Description

    Rabbit monoclonal [Y171] to Chk2 (phospho T68)
  • Host species

    Rabbit
  • Specificity

    The antibody will detect Chk2 phosphorylation on Threonine 68. This antibody may also detect all splice isoforms (2-12) based on sequence homology.
  • Tested applications

    Suitable for: IP, WB, Dot blotmore details
    Unsuitable for: Flow Cyt,ICC or IHC
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Chk2. The exact sequence is proprietary.

  • Positive control

    • Cell lysates from HEK-293 cells treated with UV and/or phosphatase.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32148 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
IP 1/20 - 1/50.
WB 1/1000 - 1/5000. Detects a band of approximately 62 kDa (predicted molecular weight: 61 kDa).
Dot blot 1/500.
  • Application notes
    Is unsuitable for Flow Cyt,ICC or IHC.
  • Target

    • Function

      Regulates cell cycle checkpoints and apoptosis in response to DNA damage, particularly to DNA double-strand breaks. Inhibits CDC25C phosphatase by phosphorylation on 'Ser-216', preventing the entry into mitosis. May also play a role in meiosis. Regulates the TP53 tumor suppressor through phosphorylation at 'Thr-18' and 'Ser-20'.
    • Tissue specificity

      High expression is found in testis, spleen, colon and peripheral blood leukocytes. Low expression is found in other tissues.
    • Involvement in disease

      Defects in CHEK2 are associated with Li-Fraumeni syndrome 2 (LFS2) [MIM:609265]; a highly penetrant familial cancer phenotype usually associated with inherited mutations in p53/TP53.
      Defects in CHEK2 may be a cause of susceptibility to prostate cancer (PC) [MIM:176807]. It is a malignancy originating in tissues of the prostate. Most prostate cancers are adenocarcinomas that develop in the acini of the prostatic ducts. Other rare histopathologic types of prostate cancer that occur in approximately 5% of patients include small cell carcinoma, mucinous carcinoma, prostatic ductal carcinoma, transitional cell carcinoma, squamous cell carcinoma, basal cell carcinoma, adenoid cystic carcinoma (basaloid), signet-ring cell carcinoma and neuroendocrine carcinoma.
      Defects in CHEK2 are found in some patients with osteogenic sarcoma (OSRC) [MIM:259500].
    • Sequence similarities

      Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CHK2 subfamily.
      Contains 1 FHA domain.
      Contains 1 protein kinase domain.
    • Post-translational
      modifications

      Phosphorylated by PLK4.
    • Cellular localization

      Nucleus; Nucleus. Isoform 10 is present throughout the cell and Nucleus > PML body. Nucleus > nucleoplasm. Recruited into PML bodies together with TP53.
    • Information by UniProt
    • Database links

    • Alternative names

      • CDS 1 antibody
      • Cds1 antibody
      • Cds1 homolog antibody
      • Checkpoint kinase 2 antibody
      • Checkpoint like protein CHK2 antibody
      • CHEK 2 antibody
      • Chek2 antibody
      • Chk 2 antibody
      • CHK2 checkpoint homolog (S. pombe) antibody
      • CHK2 checkpoint homolog antibody
      • CHK2_HUMAN antibody
      • hCds1 antibody
      • HuCds 1 antibody
      • LFS 2 antibody
      • LFS2 antibody
      • PP1425 antibody
      • RAD 53 antibody
      • RAD53 antibody
      • Rad53 homolog antibody
      • Serine/threonine protein kinase Chk2 antibody
      • Serine/threonine-protein kinase Chk2 antibody
      see all

    Images

    • ab32148 (purified) at 1:30 dilution (2µg) immunoprecipitating Chk2 in HEK-293 treated with UV whole cell lysate.
      Lane 1 (input): HEK-293 (Human embryonic kidney epithelial cell) treated with UV whole cell lysate 10µg
      Lane 2 (+): ab32148 & HEK-293 treated with UV whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32148 in HEK-293 treated with UV whole cell lysate
      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • All lanes : Anti-Chk2 (phospho T68) antibody [Y171] (ab32148) at 1/1000 dilution (Purified)

      Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysates
      Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) treated with UV at 60J/m2 for 2 hours whole cell lysates
      Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) treated with UV at 60J/m2 for 2 hours whole cell lysates, then the Membrane incubated with phosphatase

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 61 kDa
      Observed band size: 70 kDa
      why is the actual band size different from the predicted?

    • All lanes : Anti-Chk2 (phospho T68) antibody [Y171] (ab32148) at 1/200 dilution

      Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
      Lane 2 : Whole cell lysate from HEK-293 (Human epithelial cell line from embryonic kidney) cells, treated with UV at 60J/m2 for 2 hours.
      Lane 3 : Whole cell lysate from HEK-293 (Human epithelial cell line from embryonic kidney) cells, treated with UV at 60J/m2 for 2 hours. Membrane incubated with phosphatase.

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 61 kDa
      Observed band size: 70 kDa why is the actual band size different from the predicted?


      Exposure time: 10 seconds


      Blocking/dilution buffer: 5% NFDM/TBST

    • ab32148 at 1/20 dilution immunoprecipitating Chk2 (phospho T68) in HEK-293 (human embryonic kidney) whole cell lysate observed at 70kDa (lanes 1 and 2).

      Lane 1 (input): HEK-293 cells treated with UV at 60J/m2 for 2 hours. Whole cell lysate 10μg

      Lane 2 (+): ab32148 + HEK-293 treated with UV at 60J/m2 for 2 hours. Whole cell lysate

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32148 in HEK- treated with UV at 60J/m2 for 2 hours. Whole cell lysate

      For western blotting, ab32148 at 1/500 dilution was used followed by ab131366 VeriBlot for IP Detection Reagent (HRP) for detection at 1/1000.

      Blocking and Diluting buffer and concentration: 5% NFDM/TBST.

      This image was generated using the unpurified version of the product. 

    • Dot Blot analysis on immunogen phospho-peptide (A) and non-phospho peptide (B) using anti-Phospho-Chkr2 (pT68) RabMAb (ab32148) dilution 1:500

      This image was generated using the unpurified version of the product. 

    • All lanes : Anti-Chk2 (phospho T68) antibody [Y171] (ab32148) at 1/1000 dilution

      Lane 1 : HCT116 treated with DMSO (control) whole cell lysate
      Lane 2 : HCT116 treated with 10 µM etoposide for 24 hours whole cell lysate

      Lysates/proteins at 60 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 61 kDa
      Observed band size: 70 kDa why is the actual band size different from the predicted?


      Exposure time: 20 seconds


      Non-specific band : 35, 30 kDa

      Blocking/Diluting buffer : 5% Milk

       This image was generated using the unpurified version of the product. 

      See Abreview

    References

    This product has been referenced in:

    • Vincelette ND  et al. Effect of CHK1 Inhibition on CPX-351 Cytotoxicity in vitro and ex vivo. Sci Rep 9:3617 (2019). Read more (PubMed: 30837643) »
    • Sun K  et al. Oxidized ATM-mediated glycolysis enhancement in breast cancer-associated fibroblasts contributes to tumor invasion through lactate as metabolic coupling. EBioMedicine 41:370-383 (2019). Read more (PubMed: 30799198) »
    See all 10 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    Application
    Western blot
    Loading amount
    30 µg
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Sample
    Human Cell lysate - whole cell (lymphocytes)
    Specification
    lymphocytes
    Treatment
    UV treated
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 37°C

    Abcam user community

    Verified customer

    Submitted Apr 07 2014

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HCT116 colon cancer cell line)
    Loading amount
    60 µg
    Specification
    HCT116 colon cancer cell line
    Treatment
    DMSO control and 10 µM etoposide for 24 hrs
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Mr. Christian Marx

    Verified customer

    Submitted Oct 29 2012

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