STO-609

For research use only.

Catalog No.S8274

5 publications

STO-609 Chemical Structure

CAS No. 52029-86-4

STO-609 is a specific inhibitor of the Ca2+/Calmodulin-dependent protein kinase kinase(CaM-KK) that inhibits the activities of recombinant CaM-KKα and CaM-KKβ isoforms, with Ki values of 80 and 15 ng/ml, respectively, and also inhibits their autophosphorylation activities. STO-609 inhibits AMPKK activity and inhibits autophagy.

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Selleck's STO-609 has been cited by 5 publications

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Choose Selective CaMK Inhibitors

Biological Activity

Description STO-609 is a specific inhibitor of the Ca2+/Calmodulin-dependent protein kinase kinase(CaM-KK) that inhibits the activities of recombinant CaM-KKα and CaM-KKβ isoforms, with Ki values of 80 and 15 ng/ml, respectively, and also inhibits their autophosphorylation activities. STO-609 inhibits AMPKK activity and inhibits autophagy.
Targets
AMPK [4]
()
CaM-KKβ [1]
(Cell-free assay)
CaM-KKα [1]
(Cell-free assay)
47 nM(Ki) 0.25 μM(Ki)
In vitro

STO-609 inhibits the activities of recombinant CaM-KKα and CaM-KKβ isoforms, with Ki values of 80 and 15 ng/ml, respectively, and also inhibits their autophosphorylation activities. STO-609 is highly selective for CaM-KK without any significant effect on the downstream CaM kinases (CaM-KI and -IV). In transfected HeLa cells, STO-609 suppresses the Ca2+-induced activation of CaM-KIV in a dose-dependent manner. STO-609 can permeate cells and is a competitive inhibitor of ATP[1].

Assay
Methods Test Index PMID
Western blot
p-AMPK / AMPK ; 

PubMed: 27158402     


Immunoblots illustrating p-AMPK levels in gastric adenocarcinoma cells (SNU-1 and N87) treated with dimethyl sulfoxide (DMSO) (vehicle) or STO-609.

p-AKT / AKT / p-ERK / ERK; 

PubMed: 27158402     


Western blotting results show the effects of STO-609 (5 and 10 μM) in gastric adenocarcinoma cells (SNU-1 and N87) on phosphorylation of Akt and ERK.

27158402
Growth inhibition assay
Cell viability ; 

PubMed: 27158402     


Histogram illustrating the dose-dependent effect of STO-609 (2.5, 5, 10 and 20 μM) on the viability of the two gastric cancer cell lines (SNU-1 and N87).

27158402
In vivo

In vivo administration of STO-609 results in increased osteoblasts and diminished osteoclasts, conferring significant protection from ovariectomy (OVX)-induced osteoporosis in adult mice[2]. ICV administration of STO-609 in vivo did not affect the counterregulatory responses to neuroglucopenia and AMPK activation induced by glucopenia[3].

Protocol

Cell Research:

[1]

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  • Cell lines: HeLa cells
  • Concentrations: 0.01-10 μg/ml
  • Incubation Time: 6 h
  • Method:

    HeLa cells were maintained in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum. Cells were subcultured in 6-cm dishes 12 h before transfection. The cells were then transferred to serum-free medium and treated with a mixture of either 3 g of pME18s plasmid DNA or 3 g of HA(hemagglutinin-tagged)-CaM-KIV and 20 μg of LipofectAMINE reagent in 2.5 ml of medium. After 20 h of incubation, the cells were further cultured in serum-free medium for 6 h in either the absence or presence of various concentrations of STO-609 (0.01-10 μg/ml in Me2SO at a final concentration of 0.5%) and then treated with or without 1 μM ionomycin for 5 min. Stimulation was terminated by the addition of 1 ml of lysis buffer, 2 mM EDTA, 2 mM EGTA, 1% Nonidet P-40, 10% glycerol, 0.2 mM phenylmethylsulfonyl fluoride, 10 mg/liter leupeptin, 10 mg/liter trypsin inhibitor, and 1 μM microcystin LR), and the cells were lysed for 30min on ice. The cell extract was collected and centrifuged at 15,000×g for 15 min, the supernatant was precleared with 40 μl of Protein G-Sepharose for 2h at 4 °C, and the supernatant was mixed with 4 g of anti-HA antibody for 3h. 40 μl of Protein G-Sepharose was then applied to the extract and incubated overnight. The immunoprecipitated resin was washed three times with 1 ml of the lysis buffer and then washed with 1 ml of kinase buffer. Protein G-Sepharose with immunoprecipitated HA-CaM-KIV was subjected to the protein kinase assay in the presence of 1 mM EGTA using syntide-2 as a substrate.


    (Only for Reference)
Animal Research:

[2]

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  • Animal Models: WT, Camkk2−/− and Camk4−/− mice (all in C57BL/6 background)
  • Dosages: 10 μmol/kg
  • Administration: i.p.
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 8 mg/mL (25.45 mM)
Water Insoluble
Ethanol Insoluble

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 314.29
Formula

C19H10N2O3

CAS No. 52029-86-4
Storage powder
in solvent
Synonyms N/A
Smiles C1=CC=C2C(=C1)N=C3N2C(=O)C4=CC=CC5=C(C=CC3=C54)C(=O)O

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Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID