IM-12

Catalog No.S7566 Batch:S756602

Technical Data

Formula

C₂₂H₂₀FN₃O₂

Molecular Weight

377.41

CAS No.

1129669-05-1

Solubility (25°C)*

In vitro

DMSO

75 mg/mL (198.72 mM)

Ethanol

12 mg/mL (31.79 mM)

Water

Insoluble

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

Clear solution	5% DMSO 1 95% Corn oil	0.625mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 12.5 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.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O	3.75mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 75 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.

* <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

IM-12 is a selective GSK-3β inhibitor with IC50 of 53 nM, and also enhances canonical Wnt signalling.

Targets

GSK-3β ^[1]
(Cell-free assay)

53 nM

In vitro

In hNPCs, IM-12 inhibits GSK-3β activity with IC50 of 3.8 μM and subsequently increases β-catenin concentration significantly. Through the activation of canonical Wnt signalling pathway, IM-12 promotes the neuronal differentiation of human neural progenitor cells. ^[1]

Protocol (from reference)

Kinase Assay:^{^[1]}	GSK-3β activity assay IC50 of new synthesised compound IM-12 to GSK-3β is determined by a luminometric GSK-3β activity assay. Briefly, compounds are tested in different concentrations diluted in assay buffer containing final concentrations of: 4 mM MOPS pH 7.2; 0.4 mM EDTA; 1 mM EGTA; 2.5 mM β-glycerophosphate; 4 mM MgCl₂; 40 μM BSA; 0.05 mM DTT. Four microlitres of diluted compounds are added to 25 μM pGS-2 peptide substrate, 20 ng recombinant GSK-3β and 1 μM ATP to a total assay volume of 40 μl. The enzymatic reaction is stopped after 30 min of incubation at 30 °C by adding 40 μl KinaseGlo. The luminometric signal is allowed to stabilise for 10 min and the measured with a Glomax® 96 Microplate Luminometer.
Cell Assay:^{^[1]}	Cell lines ReNcell VM cells Concentrations 3 μM Incubation Time 72 hours Method To measure viable cells, 50-100 μL of cell suspension is analyzed using CASY technology with the appropriate program. ReNcell VM cells are seeded at a defined cell number and proliferated for 24 h. Then the medium is changed to proliferation medium with added substances at indicated concentrations. The cell number was determined every 24 h. Cells were exposed to the added drugs during the whole experiment, whereas the media is changed every 24 h.

Kinase Assay:^{^[1]}

GSK-3β activity assay
IC50 of new synthesised compound IM-12 to GSK-3β is determined by a luminometric GSK-3β activity assay. Briefly, compounds are tested in different concentrations diluted in assay buffer containing final concentrations of: 4 mM MOPS pH 7.2; 0.4 mM EDTA; 1 mM EGTA; 2.5 mM β-glycerophosphate; 4 mM MgCl₂; 40 μM BSA; 0.05 mM DTT. Four microlitres of diluted compounds are added to 25 μM pGS-2 peptide substrate, 20 ng recombinant GSK-3β and 1 μM ATP to a total assay volume of 40 μl. The enzymatic reaction is stopped after 30 min of incubation at 30 °C by adding 40 μl KinaseGlo. The luminometric signal is allowed to stabilise for 10 min and the measured with a Glomax® 96 Microplate Luminometer.

Cell Assay:^{^[1]}

Cell lines
ReNcell VM cells
Concentrations
3 μM
Incubation Time
72 hours
Method
To measure viable cells, 50-100 μL of cell suspension is analyzed using CASY technology with the appropriate program. ReNcell VM cells are seeded at a defined cell number and proliferated for 24 h. Then the medium is changed to proliferation medium with added substances at indicated concentrations. The cell number was determined every 24 h. Cells were exposed to the added drugs during the whole experiment, whereas the media is changed every 24 h.

References

[1] Schm?le AC, er al. Bioorg Med Chem. 2010, 18(18), 6785-6795.

Customer Product Validation

: Data from [Data independently produced by , , Exp Cell Res, 2016, 345(2):206-17.]

Selleck's IM-12 has been cited by 16 publications

Self-renewing human naïve pluripotent stem cells dedifferentiate in 3D culture and form blastoids spontaneously [ Nat Commun, 2024, 15(1):668]	PubMed: 38253551
A transient transcriptional activation governs unpolarized-to-polarized morphogenesis during embryo implantation [ Mol Cell, 2024, 84(14):2665-2681.e13]	PubMed: 38955180
Development of a live cell assay for real-time monitoring the interactions between the Hippo pathway components 14-3-3 and TAZ [ SLAS Discov, 2024, 29(8):100191]	PubMed: 39510350
Generation of a humanized mesonephros in pigs from induced pluripotent stem cells via embryo complementation [ Cell Stem Cell, 2023, 30(9):1235-1245.e6]	PubMed: 37683604
dCas9 fusion to computer-designed PRC2 inhibitor reveals functional TATA box in distal promoter region [ Cell Rep, 2022, 38(9):110457]	PubMed: 35235780
Establishment and characterization of immortalized sweat gland myoepithelial cells [ Sci Rep, 2022, 12(1):7]	PubMed: 34997030
B3GALT5 knockout alters gycosphingolipid profile and facilitates transition to human naïve pluripotency [ Proc Natl Acad Sci U S A, 2020, 117(44):27435-27444]	PubMed: 33087559
A novel human colon signet-ring cell carcinoma organoid line: establishment, characterization and application [ Carcinogenesis, 2020, 41(7):993-1004]	PubMed: 31740922
RHAMM inhibits cell migration via the AKT/GSK3β/Snail axis in luminal A subtype breast cancer [ Anat Rec (Hoboken), 2020, 303(9):2344-2356]	PubMed: 31769593
Non-canonical WNT-signaling controls differentiation of lymphatics and extension lymphangiogenesis via RAC and JNK signaling. [ Sci Rep, 2019, 9(1):4739]	PubMed: 30894622

RETURN POLICY
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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.