Overview

  • Product nameAnti-Chk2 antibody [EPR4325]
    See all Chk2 primary antibodies
  • Description
    Rabbit monoclonal [EPR4325] to Chk2
  • Tested applicationsSuitable for: ICC/IF, Flow Cyt, WB, IP, IHC-P, ICCmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant fragment corresponding to Human Chk2 aa 1-200.

  • Positive control
    • HeLa treated with Gamma Irradiation, HeLa, HT-29, and 293T cell lysates; Human colon tissue; Human spleen tissue ICC/IF: HAP1 wild-type cells and HAP1-Chk2 knockout cells
  • 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.

    Alternative versions available:

    Anti-Chk2 antibody (Alexa Fluor® 488) [EPR4325] (ab200762)
    Anti-Chk2 antibody (HRP) [EPR4325] (ab200767)

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109413 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
ICC/IF 1/250.
Flow Cyt Use at an assay dependent concentration.
WB 1/50000 - 1/200000. Detects a band of approximately 62 kDa (predicted molecular weight: 61 kDa).
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. antigen retrieval is recommended.
ICC 1/100 - 1/250.

Target

  • FunctionRegulates 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 specificityHigh expression is found in testis, spleen, colon and peripheral blood leukocytes. Low expression is found in other tissues.
  • Involvement in diseaseDefects 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 similaritiesBelongs 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 localizationNucleus; 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

Anti-Chk2 antibody [EPR4325] images



  • Predicted band size : 61 kDa

    Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 µg)
    Lanes 2, 6 and 10: Chk2 knockout HAP1 cell lysate (20 µg)
    Lanes 3, 7 and 11: HeLa cell lysate (20 µg)
    Lanes 4, 8 and 12: HEK293 cell lysate (20 µg)
    Lanes 1, 2, 3 and 4: Green signal from target -  ab109413 observed at 62 kDa
    Lanes 5, 6, 7 and 8: Red signal from loading control - ab8245 observed at 37 kDa
    Lanes 9, 10, 11 and 12: Merged (red and green) signal

    ab109413 was shown to specifically react with Chk2 when Chk2 knockout samples were used. Wild-type and Chk2 knockout samples were subjected to SDS-PAGE. ab109413 and ab8245 (loading control to GAPDH) were diluted 1/50 000 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

  • ab109413 staining Chk2 in wild-type HAP1 cells (top panel) and Chk2 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab109413 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue using 1/100 ab109413
  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Chk2 with purified ab109413 at 1/230 dilution(10ug/ml) (red). Cells were fixed with 80% methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor® 488)(ab150077)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black)(ab172730) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.



  • Predicted band size : 61 kDa

    Lanes 1: Wild-type HAP1 cell lysate (20 µg)
    Lanes 2: Chk2 knockout HAP1 cell lysate (20 µg)
    Lanes 3: HeLa cell lysate (20 µg)
    Lanes 4: HEK293 cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab109413 observed at 64 kDa. Red - loading control, ab8245, observed at 37 kDa.

    This western blot image is a comparison between ab109413 and a competitor's rabbit polyclonal antibody.

     

  • All lanes : Anti-Chk2 antibody [EPR4325] (ab109413) at 1/50000 dilution

    Lane 1 : HeLa treated with Gamma Irradiation
    Lane 2 : HeLa cell lysate
    Lane 3 : HT29 cell lysate
    Lane 4 : 293T cell lysate

    Lysates/proteins at 10 µg per lane.


    Predicted band size : 61 kDa
    Observed band size : 62 kDa (why is the actual band size different from the predicted?)
  • Immunohistochemical analysis of paraffin-embedded Human spleen tissue using 1/100 ab109413
  • ab109413 (1/500) staining Chk2 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5%Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please see Abreview.

    See Abreview

References for Anti-Chk2 antibody [EPR4325] (ab109413)

This product has been referenced in:
  • Li WF  et al. WISP-1 contributes to fractionated irradiation-induced radioresistance in esophageal carcinoma cell lines and mice. PLoS One 9:e94751 (2014). Human . Read more (PubMed: 24728101) »
  • Sowd GA  et al. SV40 utilizes ATM kinase activity to prevent non-homologous end joining of broken viral DNA replication products. PLoS Pathog 10:e1004536 (2014). Read more (PubMed: 25474690) »

See all 2 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa cell lysate)
Loading amount 22 µg
Specification HeLa cell lysate
Treatment +/- 10 Gy IR
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted May 13 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step 5% BSA + 1% NDS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Sample Human Cell (HeLa)
Specification HeLa
Permeabilization Yes - 0.1% Triton X-100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 09 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Hela, HT29)
Loading amount 30 µg
Specification Hela, HT29
Treatment see legend
Gel Running Conditions Reduced Denaturing (7% Tris acetate)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted May 02 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton X-100 in PBS
Username

Dr. Kirk Mcmanus

Verified customer

Submitted Feb 19 2013

Thank you for contacting us.

Below you can find the tested WB protocol.

Protocols: Western Blot

Cell Lysis and Western Blotting Protocol


http://www.epitomics.com/pdf/westerndata.pdf

1. Solutions ...

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