AHP1266 detects human Checkpoint kinase 2 (Chk2), phosphorylated at threonine 68 (Thr68). Chk2 is a serine/threonine-protein kinase which functions as a cell cycle checkpoint regulator. ATM and ATR kinases are activated in response to ionizing and ultraviolet radiation, and phsophorylate Thr68 and other sites in the amino-terminal region of Chk2, leading to Chk2 dimerisation and activation.
Activated Chk2 phosphorylates and inhibits Cdc25C phosphatase, preventing the entry into mitosis, and has been shown to stabilize the tumour suppressor protein p53, leading to cell cycle arrest in G1. In addition, Chk2 can phosphorylate BRCA1, allowing BRCA1 to restore survival after DNA damage.
Chk2 is a putative tumour suppressor protein, with mutations in to the Chk2 gene linked to Li-Fraumeni syndrome, a familial cancer usually associated with inherited mutations in TP53.
Species Cross-Reactivity
Target Species
Cross Reactivity
Chimpanzee
Expected from Sequence
Mouse
Expected from Sequence
Rat
Expected from Sequence
Application
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visitwww.abdserotec.com/protocols.
Application Name
Yes
No
Not Determined
Suggested Dilution
ELISA
1/10000 - 1/50000
Flow Cytometry
Functional Assays
Immunohistology - Frozen
Immunohistology - Paraffin
Immunoprecipitation
Western Blotting(1)
1/200 - 1/2000
(1)
For the detection of phosphoproteins, tyrosine phosphatase inhibitors such as 1mM Sodium Orthovanadate should be added to the sample buffer. Milk or other casein-based blocking solutions are not recommended as casein is a phosphoprotein and its use can result in high background.
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Western Blotting
AHP1266 detects a band of approximately 61kDa.
Storage
Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
18 months from date of despatch.
Antiserum Preparation
Antisera to Chk2 (pTHR68) were raised by repeated immunisations of rabbits with highly purified antigen. Purified IgG prepared by affinity chromatography.
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