Anti-Chk2 (phospho T68) antibody (ab85743)
Key features and details
- Rabbit polyclonal to Chk2 (phospho T68)
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Chk2 (phospho T68) antibody
See all Chk2 primary antibodies -
Description
Rabbit polyclonal to Chk2 (phospho T68) -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Cow, Pig
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- This antibody produced a positive signal in Irradiated HeLa. It also gave a positive result in FFPE human colon adenocarcinoma tissue sections.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 1% BSA
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
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.
Application Abreviews Notes 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). Target
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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
modificationsPhosphorylated 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
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Database links
- Entrez Gene: 518897 Cow
- Entrez Gene: 11200 Human
- Entrez Gene: 50883 Mouse
- Entrez Gene: 100192447 Pig
- Entrez Gene: 114212 Rat
- Omim: 604373 Human
- SwissProt: O96017 Human
- SwissProt: Q9Z265 Mouse
see all -
Alternative names
- CDS 1 antibody
- Cds1 antibody
- Cds1 homolog antibody
see all
Images
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Western blot - Anti-Chk2 (phospho T68) antibody (ab85743)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.
Secondary
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 minutesSecondary antibody - anti-rabbit HRP (IgG H&L; ab97080)
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Immunocytochemistry/ Immunofluorescence - Anti-Chk2 (phospho T68) antibody (ab85743)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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chk2 (phospho T68) antibody (ab85743)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.
Protocols
Datasheets and documents
References (5)
ab85743 has been referenced in 5 publications.
- Saxena S et al. ATR Signaling Uncouples the Role of RAD51 Paralogs in Homologous Recombination and Replication Stress Response. Cell Rep 29:551-559.e4 (2019). PubMed: 31618626
- 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
Images
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Western blot - Anti-Chk2 (phospho T68) antibody (ab85743)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.
Secondary
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 minutesSecondary antibody - anti-rabbit HRP (IgG H&L; ab97080)
-
Immunocytochemistry/ Immunofluorescence - Anti-Chk2 (phospho T68) antibody (ab85743)
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.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chk2 (phospho T68) antibody (ab85743)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.
