PluriSIn #1 (NSC 14613)

Catalog No.S8076 Batch:S807601

Technical Data

Formula

C₁₂H₁₁N₃O

Molecular Weight

213.24

CAS No.

91396-88-2

Solubility (25°C)*

In vitro

DMSO

43 mg/mL (201.65 mM)

Water

Insoluble

Ethanol

Insoluble

In vivo (Add solvents to the product individually and in order)

Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	2.15mg/ml (10.08mM)	Taking the 1 mL working solution as an example, add 50 μL of 43 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	5% DMSO 1 95% Corn oil Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	0.31mg/ml (1.45mM)	Taking the 1 mL working solution as an example, add 50 μL of 6.2 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.

* <1 mg/ml means slightly soluble or 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.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description

PluriSIn #1 (NSC 14613) is an inhibitor of the stearoyl-coA desaturase 1 (SCD1), which is able to selectively eliminate hPSCs.

Targets

SCD1 ^[1]

In vitro

PluriSIn #1 (NSC 14613) has a robust, rapid, and selective cytotoxic effect toward hPSCs, with apoptosis as the central cell death mechanism it activates. This compound leads to ER stress in hPSCs, induces a ~30% decrease in protein synthesis at 20 μM, and causes a ~65% decrease in stearoyl-coA desaturase (SCD1) activity. It also prevents teratoma formation from undifferentiated hPSCs at 20 μM. PluriSIns inhibit mPSCs and hinder 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 of PluriSIn #1 (NSC 14613) 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 Assay:^{^[1]}	Cell lines Human iPSC line BJ-iPS28 Concentrations ~20 μM Incubation Time 1 days Method PluriSIn #1 (NSC 14613) 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.

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 of PluriSIn #1 (NSC 14613) 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 Assay:^{^[1]}

Cell lines
Human iPSC line BJ-iPS28
Concentrations
~20 μM
Incubation Time
1 days
Method
PluriSIn #1 (NSC 14613) 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.

References

https://pubmed.ncbi.nlm.nih.gov/23318055/

Selleck's PluriSIn #1 (NSC 14613) Has Been Cited by 4 Publications

CTC-derived pancreatic cancer models serve as research tools and are suitable for precision medicine approaches [ Cell Rep Med, 2024, 5(9):101692]	PubMed: 39163864
Targeting a Lipid Desaturation Enzyme, SCD1, Selectively Eliminates Colon Cancer Stem Cells through the Suppression of Wnt and NOTCH Signaling [ Cells, 2021, 10(1)E106]	PubMed: 33430034
Clinical and biochemical relevance of monounsaturated fatty acid metabolism targeting strategy for cancer stem cell elimination in colon cancer. [ Biochem Biophys Res Commun, 2019, 519(1):100-105]	PubMed: 31481234
Clinical and biochemical relevance of monounsaturated fatty acid metabolism targeting strategy for cancer stem cell elimination in colon cancer [ Biochem Biophys Res Commun, 2019, 519(1):100-105]	PubMed: 31481234

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SHIPPING AND STORAGE
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