Key features and details
- Rabbit polyclonal to Chk2 (phospho T68)
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Human
- Isotype: IgG
Product nameAnti-Chk2 (phospho T68) antibody
See all Chk2 primary antibodies
DescriptionRabbit polyclonal to Chk2 (phospho T68)
Tested applicationsSuitable for: ICC/IF, IHC-P, WBmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat, Cow, Pig
Synthetic peptide corresponding to Human Chk2 aa 50-150 conjugated to keyhole limpet haemocyanin.
(Peptide available as
- This antibody produced a positive signal in Irradiated HeLa. It also gave a positive result in FFPE human colon adenocarcinoma tissue sections.
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
Constituents: PBS, 1% BSA
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
Our Abpromise guarantee covers the use of ab85743 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 70 kDa (predicted molecular weight: 61 kDa).|
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
All lanes : Anti-Chk2 (phospho T68) antibody (ab85743) at 1 µg/ml
Lane 1 : HeLa Whole Cell Lysate - Irradiated (5Gy)
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : HeLa Whole Cell Lysate - Irradiated (5Gy) with Human Chk2 (phospho T68) peptide (ab111650) at 1 µg/ml
Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human Chk2 (phospho T68) peptide (ab111650) at 1 µg/ml
Lane 5 : HeLa Whole Cell Lysate - Irradiated (5Gy) with Human Chk2 peptide (ab118179) at 1 µg/ml
Lane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human Chk2 peptide (ab118179) at 1 µg/ml
Lysates/proteins at 10 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 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?
Additional bands at: 145 kDa, 38 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 3 minutes
ICC/IF image of ab85743 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab85743 at 5µg/ml overnight at +4°C. The secondary antibody (green) was goat anti rabbit DyLight® 488 (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in formaldehyde fixed (4%, 10min) HeLa cells at 5ug/ml.
IHC image of Chk2 staining in human colon adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab85743, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab85743 has been referenced in 4 publications.
- Huang D et al. Isoorientin triggers apoptosis of hepatoblastoma by inducing DNA double-strand breaks and suppressing homologous recombination repair. Biomed Pharmacother 101:719-728 (2018). WB ; Human . PubMed: 29524880
- Saxena S et al. XRCC2 Regulates Replication Fork Progression during dNTP Alterations. Cell Rep 25:3273-3282.e6 (2018). PubMed: 30566856
- Wang H et al. Azidothymidine inhibits cell growth and telomerase activity and induces DNA damage in human esophageal cancer. Mol Med Rep 15:4055-4060 (2017). PubMed: 28487971
- Lagunas AM et al. Telomere DNA damage signaling regulates cancer stem cell evolution, epithelial mesenchymal transition, and metastasis. Oncotarget 8:80139-80155 (2017). PubMed: 29113290