Product nameAnti-Chk2 antibody [EPR4325]
See all Chk2 primary antibodies
DescriptionRabbit monoclonal [EPR4325] to Chk2
Tested applicationsSuitable for: ICC/IF, Flow Cyt, WB, IP, IHC-P, ICCmore details
Species reactivityReacts with: Human
Recombinant fragment within Human Chk2 aa 1-200. The exact sequence is proprietary.
Database link: O96017
- WB: HeLa (untreated and treated with gamma irradiation), HT-29, and 293T cell lysates. IHC-P: Human colon and spleen tissues. ICC/IF: Wild-type HAP1 cells.
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
This product is a recombinant rabbit monoclonal antibody.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
PurityTissue culture supernatant
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.
|Flow Cyt||Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|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.|
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.
modificationsPhosphorylated 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
- CDS 1 antibody
- Cds1 antibody
- Cds1 homolog antibody
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 (5 minutes), 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 1 hour. 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 1 hour 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 ab109413 at a 1/100 dilution.
Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Chk2 with purified ab109413 at 1/230 dilution(10 µg/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.
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 (human epithelial cell line from cervix adenocarcinoma) cells treated with gamma irradiation
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 3 : HT-29 (human colorectal adenocarcinoma cell line) cell lysate
Lane 4 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) 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 ab109413 at a 1/100 dilution.
ab109413 (1/500) staining Chk2 in HeLa (human epithelial cell line from cervix adenocarcinoma) cells (green). Cells were fixed in paraformaldehyde, permeabilized 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.
This product has been referenced in:
- Yang F et al. HPRT1 activity loss is associated with resistance to thiopurine in ALL. Oncotarget 9:2268-2278 (2018). Read more (PubMed: 29416770) »
- Zhu M et al. Expression of DNA doublestrand repair proteins in oral leukoplakia and the risk of malignant transformation. Oncol Lett 15:9827-9835 (2018). Read more (PubMed: 29928356) »