SC79

Catalog No.S7863

SC79 Chemical Structure

Molecular Weight(MW): 364.78

SC79 is a brain-penetrable Akt phosphorylation activator and an inhibitor of Akt-PH domain translocation.

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  • The molecular structure along with the molecular weight (MW) of SC79 were presented (A). ARPE-19 cells were treated with applied concentration of SC79 (0.1–10 μg/mL, or 2.74–27.41 μM) for 1 h, p-Akt (Ser-473) and Akt1 expression was tested by Western blots and was quantified (B) n = 4). ARPE-19 cells (C and D) primary murine RPE cells ("Primary RPE", (E)) or HLECs (F) pretreated with applied concentration of SC79 for 30 min, were subjected to UV radiation (30 mJ/cm2), cells were further cultured for 24 h, and cell viability was tested by MTT assay (C, E and F); Cell death was detected by trypan blue staining assay (D). "Ctrl" stands for untreated control group (Same for all figures). For each assay, n = 5. Experiments in this figure were repeated three times to insure consistency of results. *p < 0.05 vs. “Ctrl” group (C–F). **p < 0.05 vs. UV only group (C-F).

    Oncotarget, 2016, 7(37):60123-60132. SC79 purchased from Selleck.

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Biological Activity

Description SC79 is a brain-penetrable Akt phosphorylation activator and an inhibitor of Akt-PH domain translocation.
Targets
Akt [1]
In vitro

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. [1] SC79 restores proliferation of BRAT1 knockdown cells, and reduces the production of superoxide in mitochondria of MitoSox positive cells. [2]

In vivo In the permanent focal cerebral ischemia mouse model, SC79 (0.04 mg/g, i.p.) enables the cytosolic activation of Akt, and recapitulates the primary cellular function of Akt signaling, resulting in augmented neuronal survival. [1]

Protocol

Kinase Assay:

[1]

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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 Research:

[1]

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  • Cell lines: HsSultan and NB4 cells
  • Concentrations: 8 μg/mL
  • Incubation Time: 24 h
  • Method:

    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.


    (Only for Reference)
Animal Research:

[1]

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  • Animal Models: Permanent focal cerebral ischemia mouse model
  • Formulation: DMSO
  • Dosages: 0.04 mg/g
  • Administration: i.p.
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 72 mg/mL (197.37 mM)
Ethanol 72 mg/mL warmed (197.37 mM)
Water Insoluble
In vivo Add solvents individually and in order:
2% DMSO+corn oil
5mg/mL

* 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 364.78
Formula

C17H17ClN2O5

CAS No. 305834-79-1
Storage powder
Synonyms N/A

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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