Overview

  • Product name

    Anti-Chk2 antibody [EPR19482]
    See all Chk2 primary antibodies
  • Description

    Rabbit monoclonal [EPR19482] to Chk2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human Chk2 aa 50-300. The exact sequence is proprietary.
    Database link: O96017

  • Positive control

    • WB: HeLa, HEK-293, HEK-293T, HCT 116 and SH-SY5Y whole cell lysates; human colon, thymus and kidney lysates. IHC-P: Human testis and breast cancer tissues. ICC/IF: HeLa and HCT 116 cells. Flow Cyt: HCT 116 cells. IP: HeLa and HEK-293T whole cell lysates.
  • General notes

     

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab207446 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 1/1000. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/500.
Flow Cyt 1/70.
IP 1/40.

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

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: CHEK2 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)

    Lanes 1 - 3: Merged signal (red and green). Green - ab207446 observed at 68 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab207446 was shown to specifically recognize CHEK2 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when CHEK2 knockout samples were examined. Wild-type and CHEK2 knockout samples were subjected to SDS-PAGE. Ab207446 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Lanes 1-3 & 5 : Anti-Chk2 antibody [EPR19482] (ab207446) at 1/1000 dilution
    Lane 4 : Anti-Chk2 antibody [EPR19482] (ab207446) at 1/5000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 3 : HEK-293T (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 4 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
    Lane 5 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 61 kDa
    Observed band size: 61 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/3: 15 seconds; Lane 2: 8 seconds; Lane 4: 30 seconds; Lane 5: 3 minutes.

  • Immunohistochemical analysis of paraffin-embedded Hhman testis tissue labeling Chk2 with ab207446 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human spermatogonium is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • All lanes : Anti-Chk2 antibody [EPR19482] (ab207446) at 1/1000 dilution

    Lane 1 : Human colon lysate
    Lane 2 : Human thymus lysate
    Lane 3 : Human kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 61 kDa
    Observed band size: 61 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Chk2 with ab207446 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:

    -ve control 1: ab207446 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution.

    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Chk2 with ab207446 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on tumor cells of human breast cancer is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (Human colorectal carcinoma cell line) cells labeling Chk2 with ab207446 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HCT 116 cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab207446 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed HCT 116 (Human colorectal carcinoma cell line) cells labeling Chk2 with ab207446 at 1/70 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

    Note: Cells were permeabilised with 90% methanol-PBS, -20°C, 30min

  • Chk2 was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab207446 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab207446 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate 10µg (Input).

    Lane 2: ab207446 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207446 in HeLa whole cell lysate.

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

    Exposure time: 10 seconds.

  • Chk2 was immunoprecipitated from 1mg of HEK-293T (Human epithelial cell line from embryonic kidney) whole cell lysate with ab207446 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab207446 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HEK-293T whole cell lysate 10µg (Input).

    Lane 2: ab207446 IP in HEK-293T whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207446 in HEK-293T whole cell lysate.

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

    Exposure time: 10 seconds.

References

ab207446 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS cells)
Gel Running Conditions
Reduced Denaturing (4-12% Gradient Gel)
Loading amount
50 µg
Specification
U2OS cells
Blocking step
Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C

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Submitted Oct 11 2016

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