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KU-55933

Catalog No. T2685 CAS 587871-26-9

Synonyms: ATM Kinase Inhibitor, KU55933, Inhibitor, inhibit, Ataxia telangiectasia mutated, KU-55933, KU 55933, ATM/ATR, ATM and RAD3 related, Autophagy

KU-55933 (ATM Kinase Inhibitor) is a potent and specific ATM inhibitor.

All products from TargetMol are for Research Use Only. Not for Human or Veterinary or Therapeutic Use.

KU-55933, CAS 587871-26-9

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Purity: 98%

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Description	KU-55933 (ATM Kinase Inhibitor) is a potent and specific ATM inhibitor.
Targets&IC50	ATM:12.9 nM
Kinase Assay	Purified enzyme assays: ATM for use in the in vitro assay is obtained from HeLa nuclear extract by immunoprecipitation with rabbit polyclonal antiserum raised to the COOH-terminal 400 amino acids of ATM in buffer containing 25 mM HEPES (pH 7.4), 2 mM MgCl2, 250 mM KCl, 500 μM EDTA, 100 μM Na3VO4, 10% v/v glycerol, and 0.1% v/v Igepal. ATM-antibody complexes are isolated from nuclear extract by incubating with protein A-Sepharose beads for 1 hour and then through centrifugation to recover the beads. In the well of a 96-well plate, ATM-containing Sepharose beads are incubated with 1 μg of substrate glutathione S-transferase–p53N66 (NH2-terminal 66 amino acids of p53 fused to glutathione S-transferase) in ATM assay buffer [25 mM HEPES (pH 7.4), 75 mM NaCl, 3 mM MgCl2, 2 mM MnCl2, 50 μM Na3VO4, 500 μM DTT, and 5% v/v glycerol] at 37 °C in the presence or absence of inhibitor. After 10 minutes with gentle shaking, ATP is added to a final concentration of 50 μM and the reaction continued at 37 °C for an additional 1 hour. The plate is centrifuged at 250 × g for 10 minutes (4 °C) to remove the ATM-containing beads, and the supernatant is removed and transferred to a white opaque 96-well plate and incubated at room temperature for 1.5 hours to allow glutathione S-transferase-p53N66 binding. This plate is then washed with PBS, blotted dry, and analyzed by a standard ELISA technique with a phospho-serine 15 p53 antibody. The detection of phosphorylated glutathione S-transferase-p53N66 substrate is performed in combination with a goat antimouse horseradish peroxidase-conjugated secondary antibody. Enhanced chemiluminescence solution is used to produce a signal and chemiluminescent detection is carried out.
Cell Research	U2OS cells are exposed to ionizing radiation (3, 5, or 15 Gy) or UV (5 or 50 J/m2) and the ATM response determined by Western blot analysis of p53 serine 15 phosphorylation and stabilization of wild-type p53. Whole cell extracts are obtained from each time point, proteins separated by SDS-PAGE, and the ATM-specific increase in phosphorylated serine 15 measured with a p53 phospho-serine 15 specific antibody. Overall p53 stabilization with time is also observed with a p53-specific antibody (DO-1). Similarly, for studying ATM-dependent phosphorylations on H2AX, CHK1, NBS1, and SMC1, the following antibodies are used: CHK1 phospho-serine 345 and NBS1 phospho-serine 343 antibodies. Histone H2A (H-124) and CHK1 antibodies are also used, as well as SMC1 and SMC1 phospho-serine 966 antibodies. For determination of a cellular IC50 for KU-55933, the peak response time for p53 serine 15 phosphorylation of 2 hours is used to monitor inhibition of ATM. KU-55933 is titrated onto cells and preincubated for 1 hour before ionizing radiation. Using scanning densitometry, the percentage inhibition relative to vehicle control is calculated, and the IC50 value is calculated as for the in vitro determinations.(Only for Reference)

Synonyms	ATM Kinase Inhibitor, KU55933, Inhibitor, inhibit, Ataxia telangiectasia mutated, KU-55933, KU 55933, ATM/ATR, ATM and RAD3 related, Autophagy
Molecular Weight	395.49
Formula	C21H17NO3S2
CAS No.	587871-26-9

Storage

Powder: -20°C for 3 years

In solvent: -80°C for 2 years

Solubility Information

Ethanol: 19.8 mg/mL (50 mM)

DMSO: 39.6 mg/mL (100 mM)

( < 1 mg/ml refers to the product slightly soluble or insoluble )

Citations

References and Literature

1. Huang C Y, Hsieh F S, Wang C Y, et al. Supplementary Methods Palbociclib enhances radiosensitivity of hepatocellular carcinoma and cholangiocarcinoma via inhibiting ATM-mediated DNA damage response. EUROPEAN JOURNAL OF CANCER. 2019 1. Hickson I, et al. Cancer Res. 2004, 64(24), 9152-9159. 2. Hu L, Li B, Chen G, et al A novel M phase blocker, DCZ3301 enhances the sensitivity of bortezomib in resistant multiple myeloma through DNA damage and mitotic catastrophe. Journal of Experimental & Clinical Cancer Research. 2020, 39(1): 1-14 2. Soleimani R, et al. Aging. 2011, 3(8), 782-793. 3. Ivanov VN, et al. Cancer Res. 2009, 69(8), 3510-3519. 4. Hu L, Li B, Chen G, et al. A novel M phase blocker, DCZ3301 enhances the sensitivity of bortezomib in resistant multiple myeloma through DNA damage and mitotic catastrophe. Journal of Experimental & Clinical Cancer Research. 2020, 39(1): 1-14. 5. Huang C Y, Hsieh F S, Wang C Y, et al. Supplementary Methods Palbociclib enhances radiosensitivity of hepatocellular carcinoma and cholangiocarcinoma via inhibiting ATM-mediated DNA damage response. EUROPEAN JOURNAL OF CANCER.

Related compound libraries

This product is contained In the following compound libraries：

Anti-Cancer Compound Library Preclinical Compound Library Autophagy Compound Library Anti-Lung Cancer Compound Library HIF-1 Signaling Pathway Compound Library Cancer Cell Differentiation Compound Library Glycometabolism Compound Library Anti-Cancer Metabolism Compound Library Anti-Obesity Compound Library Glutamine Metabolism Compound Library

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