ATM (ataxia telangiectasia mutated) 분석 서비스


개발하신 lead compounds에 대한 ATM (ataxia telangiectasia mutated) activity를 ELISA 방법으로 측정해 드립니다.
		Eurofins

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특징

ATM (ataxia telangiectasia mutated) 은 phosphatidylinositol 3-kinase related kinase (PIKK) family에 속하는 serine/threonine kinase 입니다. 이 kinases 그룹은 mTOR, ATR, DNA-PK를 포함합니다.
Cell cycle arrest의 결과로써, DNA 손상과 관련된 ATM 단백질의 활성을 억제하는 inhibitor를 분석합니다.
세포 내에서 ATM에 대한 타겟 substrate 중 하나인 p53의 인산화를 모니터링함으로써, ATM activity를 측정합니다.
분석 서비스는 한달에 한번 진행되며, 결과 발송까지 5-6주 소요됩니다.

원리

ATM activity can be detected by ELISA. Following the incubation with c-Myc-tagged p53 and ATP, the products are captured onto a microplate coated with anti-c-Myc antibody. The reaction components are washed away leaving just the p53 on the plate. Detection of phosphorylated p53 is achieved using a phospho-specific antibody to Ser-15 of p53.

Figure 1. ELISA signal is specific to purified ATM. ATM was titrated to demonstrate p53 phosphorylation activity. Negative controls were derived from cells transfected with empty vector, or with a kinase which does not phosphorylate Ser-15 of p53.

Figure 2. Western blot also demonstrates p53 phosphorylation at Ser-15. Left hand panel is the ponceau stain of the nitrocellulose membrane before blotting. Right hand panel is the western blot. Lanes (L-R) are: markers; ATM + p53 (no ATP); ATM + ATP (no p53); ATM + ATP + p53; p53 only.

Figure 3. The ATM assay can be used to screen inhibitors. ATM was assayed by ELISA in the presence of selected small molecule inhibitors at 1 and 10 μM.

Figure 4. Dose responses for selected inhibitors. EC50s are: KU-55933, 7 nM; PI-103, 450 nM, PIK-90, 500 nM.

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