PluriSIn #1 (NSC 14613)

Catalog No.S8076

For research use only.

PluriSIn #1 is an inhibitor of the stearoyl-coA desaturase 1 (SCD1), which is able to selectively eliminate hPSCs.

PluriSIn #1 (NSC 14613) Chemical Structure

CAS No. 91396-88-2

Selleck's PluriSIn #1 (NSC 14613) has been cited by 2 Publications

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Biological Activity

Description PluriSIn #1 is an inhibitor of the stearoyl-coA desaturase 1 (SCD1), which is able to selectively eliminate hPSCs.
Targets
SCD1 [1]
In vitro

PluriSIns #1 has a robust, rapid, and selective cytotoxic effect toward hPSCs. Apoptosis is the central cell death mechanism activated by PluriSIn #1. PluriSIn #1 leads to ER stress in hPSCs. PluriSIn #1 (20 μM) induces ~30% decrease in protein synthesis in hPSCs. PluriSIn #1 exposure induces a ~65% decrease in stearoyl-coA desaturase (SCD1) activity in hPSCs. PluriSIn #1 (20 μM) prevents teratoma gormation from undifferentiated hPSCs PluriSIns also inhibits mPSCs and hinders mouse embryonic development. [1]

Protocol (from reference)

Kinase Assay:[1]
  • SCD1 activity assays:

    Cells are plated in 6-well plates at a density of 50k to 100k cells per well. 24 h later, 20 μM PluriSIn #1 or 0.2% DMSO-control are added to the cells. After 12 h of incubation at 37 ℃, 5% CO2, the old medium is removed, cells are washed with PBS, and new medium containing 2.3 μM of 0.75 UCi [1-14C] Stearic Acid is added. The cells are incubated for up to 4 h at 37 ℃, 5% CO2. After the incubation period, the medium is discarded and the cells are washed 3 times with 2 mL of PBS. 2 mL of the mixture n-hexane: isopropanol (3:2 v:v) are added, and the cells are incubated for 30 min at 37 ℃, 5% CO2. 2 mL Folch solution (chloroform: methanol,2:1,v:v) are subsequently added. The liquid is transferred to tubes for phase partition by adding 1 mL water. The lower organic phase is evaporated and used for lipid saponification and TLC separation of the free [1-14C] Stearic Acid (substrate) and [1-14C] Oleic Acid (formed product). Lipids extracted from the cells are applied to TLC plates previously immersed in 10% NO3 Ag and activated at 120℃x60 min. Unlabeled stearic and oleic acid are added to each application point as carriers and as internal standards for identification. The plates are run with a solvent mixture of Chloroform:MeOH:AcH:DDW (90:8:1:0.8). The free fatty acids are detected by U.V. after spraying the TLC with a 2',7',dichlorofluorescein solution. The spots corresponding to stearic and oleic acid are scraped and the radioactivity counted in a sc intillating counter. SCD1 desaturase activity is calculated from the percent conversion of substrate to product and the conversion to pmol/min/106 cells.

Cell Research:[1]
  • Cell lines: Human iPSC line BJ-iPS28
  • Concentrations: ~20 μM
  • Incubation Time: 1 days
  • Method: Relative cell numbers are determined by fixating the cells with 0.5% glutardialdehyde and staining with methylene blue dissolved in 0.1 M boric acid (pH 8.5). Color extraction is performed using 0.1 M hydrochloric acid, and the staining (which is proportional to cell number) is quantitated by measuring absorbance at 650 nM.

Solubility (25°C)

In vitro

Chemical Information

Molecular Weight 213.24
Formula

C12H11N3O

CAS No. 91396-88-2
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles C1=CC=C(C=C1)NNC(=O)C2=CC=NC=C2

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Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

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Tech Support

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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