product name SC 79
Description: SC79 is a brain-penetrable Akt phosphorylation activator and an inhibitor of Akt-PH domain translocation. SC79 binds to the pleckstrin homology domain of Akt and enhances Akt phosphorylation by upstream protein kinases; SC79 also enables cytosolic activation of Akt. SC79 was shown to suppress excitotoxicity-induced neuronal death in vitro and in vivo. It exhibits brain penetrance. SC79in the cytosol, specifically binding to the PH domain of Akt. The conformation of SC79-bound Akt is favorable for phosphorylation by upstream protein kinases.
References: Proc Natl Acad Sci U S A. 2012 Jun 26;109(26):10581-6.
364.78
Formula
C17H17ClN2O5
CAS No.
305834-79-1
Storage
-20℃ for 3 years in powder form
-80℃ for 2 years in solvent
Solubility (In vitro)
DMSO: 72 mg/mL (197.4 mM)
Water: <1 mg/mL
Ethanol: 72 mg/mL (197.4 mM)
Solubility (In vivo)
2% DMSO+corn oil: 5mg/mL
Chemical Name
Ethyl 2-amino-6-chloro-4-(1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate
other peoduct :References PubMed ID：:http://www.ncbi.nlm.nih.gov/pubmed/19394490
| In Vitro |
Kinase Assay: Cytosolic phosphorylation of Akt: Hela cells are serum starved for 1 hr and treated with IGF (100ng/mL) or SC79 (4 μg/mL) for 30 minutes. Cells are lysed in Lysis buffer containing 250 mM Sucrose, 20 mM HEPES, 10 mM KCl, 1.5 mM MgCl2, 1 mM EDTA, 1 mM EGTA supplemented with protease inhibitors. Cells are passed through 25G needle several times and kept on ice for 20 minutes. Total cell lysate is taken at this point. Cell lysates are centrifuged at 100,000g for 30 minutes. Supernatant is collected as the cytosolic fraction. Pellet is washed with lysis buffer and represents the membrane fraction. Total cell lysate, cytosolic and membrane fractions are resolved by SDS-PAGE and analyzed for phospho-Akt (S473), Total Akt, Tubulin (cytosolic marker) and Orai1 (membrane marker) by western blotting. Cell Assay: HsSultan or NB4 cells (2.5 × 105) are plated in a 24-well plate in 500 μL of phenol red-free RPMI medium supplemented with 10% FBS. After incubation for 24 hours, each compound (8 µg/mL) is added and cultured for overnight (16–20 h). Fifty microliters of MTT solution (5 mg/mL in PBS) are added to each well. Following 2 hrs incubation, the purple formazan crystals are dissolved by directly adding in 500 μL of isopropanol with 0.1 M HCl to each well. After clearing the cell debris by centrifugation, the absorbance is measured at a wavelength of 570 nm. SC79 suppresses PHAKTM-GFP plasma membrane translocation, and enhances phosphorylation of all three Akt isoforms in HEK293, HeLa, HL60, NB4, and HsSulton (B cells) cells. SC79 reduces neuronal excitotoxicity and prevents stroke-induced neuronal death. SC79 restores proliferation of BRAT1 knockdown cells, and reduces the production of superoxide in mitochondria of MitoSox positive cells. |
|---|---|
| In Vivo | In the permanent focal cerebral ischemia mouse model, SC79 (0.04 mg/g, i.p.) inhibits the cytosolic activation of Akt, and recapitulates the primary cellular function of Akt signaling, resulting in augmented neuronal survival. |
| Animal model | Permanent focal cerebral ischemia mouse model |
| Formulation & Dosage | Dissolved in DMSO; 40 mg/kg; p.o. |
| References | [1] Jo H, et al. Proc Natl Acad Sci U S A. 2012, 109(26), 10581-10586. |
Related Posts
product name SC 79
Description: SC79 is a brain-penetrable Akt phosphorylation activator and an inhibitor of Akt-PH domain translocation. SC79 binds to the pleckstrin homology domain of Akt and enhances Akt phosphorylation by upstream protein kinases; SC79 also enables cytosolic activation of Akt. SC79 was shown to suppress excitotoxicity-induced neuronal death in vitro and in vivo. It exhibits brain penetrance. SC79in the cytosol, specifically binding to the PH domain of Akt. The conformation of SC79-bound Akt is favorable for phosphorylation by upstream protein kinases.
References: Proc Natl Acad Sci U S A. 2012 Jun 26;109(26):10581-6.
364.78
Formula
C17H17ClN2O5
CAS No.
305834-79-1
Storage
-20℃ for 3 years in powder form
-80℃ for 2 years in solvent
Solubility (In vitro)
DMSO: 72 mg/mL (197.4 mM)
Water: <1 mg/mL
Ethanol: 72 mg/mL (197.4 mM)
Solubility (In vivo)
2% DMSO+corn oil: 5mg/mL
Chemical Name
Ethyl 2-amino-6-chloro-4-(1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate
other peoduct :References PubMed ID：:http://www.ncbi.nlm.nih.gov/pubmed/19394490
| In Vitro |
Kinase Assay: Cytosolic phosphorylation of Akt: Hela cells are serum starved for 1 hr and treated with IGF (100ng/mL) or SC79 (4 μg/mL) for 30 minutes. Cells are lysed in Lysis buffer containing 250 mM Sucrose, 20 mM HEPES, 10 mM KCl, 1.5 mM MgCl2, 1 mM EDTA, 1 mM EGTA supplemented with protease inhibitors. Cells are passed through 25G needle several times and kept on ice for 20 minutes. Total cell lysate is taken at this point. Cell lysates are centrifuged at 100,000g for 30 minutes. Supernatant is collected as the cytosolic fraction. Pellet is washed with lysis buffer and represents the membrane fraction. Total cell lysate, cytosolic and membrane fractions are resolved by SDS-PAGE and analyzed for phospho-Akt (S473), Total Akt, Tubulin (cytosolic marker) and Orai1 (membrane marker) by western blotting. Cell Assay: HsSultan or NB4 cells (2.5 × 105) are plated in a 24-well plate in 500 μL of phenol red-free RPMI medium supplemented with 10% FBS. After incubation for 24 hours, each compound (8 µg/mL) is added and cultured for overnight (16–20 h). Fifty microliters of MTT solution (5 mg/mL in PBS) are added to each well. Following 2 hrs incubation, the purple formazan crystals are dissolved by directly adding in 500 μL of isopropanol with 0.1 M HCl to each well. After clearing the cell debris by centrifugation, the absorbance is measured at a wavelength of 570 nm. SC79 suppresses PHAKTM-GFP plasma membrane translocation, and enhances phosphorylation of all three Akt isoforms in HEK293, HeLa, HL60, NB4, and HsSulton (B cells) cells. SC79 reduces neuronal excitotoxicity and prevents stroke-induced neuronal death. SC79 restores proliferation of BRAT1 knockdown cells, and reduces the production of superoxide in mitochondria of MitoSox positive cells. |
|---|---|
| In Vivo | In the permanent focal cerebral ischemia mouse model, SC79 (0.04 mg/g, i.p.) inhibits the cytosolic activation of Akt, and recapitulates the primary cellular function of Akt signaling, resulting in augmented neuronal survival. |
| Animal model | Permanent focal cerebral ischemia mouse model |
| Formulation & Dosage | Dissolved in DMSO; 40 mg/kg; p.o. |
| References | [1] Jo H, et al. Proc Natl Acad Sci U S A. 2012, 109(26), 10581-10586. |