CHIR-98014

Catalog No.S2745 Batch:S274503

Technical Data

Formula

C₂₀H₁₇Cl₂N₉O₂

Molecular Weight

486.31

CAS No.

252935-94-7

Solubility (25°C)*

In vitro

DMSO

10 mg/mL (20.56 mM)

Water

Insoluble

Ethanol

Insoluble

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

Clear solution

10%DMSO 1 90%Corn oil

1.1mg/ml

Taking the 1 mL working solution as an example, add 100 μL of 11 mg/ml clear DMSO stock solution to 900 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

CHIR-98014 (CT98014) is a potent GSK-3α/β inhibitor with IC50 of 0.65 nM/0.58 nM in cell-free assays, with the ability to distinguish GSK-3 from its closest homologs Cdc2 and ERK2.

Targets

GSK-3β ^[1] (Cell-free assay)	GSK-3α ^[1] (Cell-free assay)
0.58 nM	0.65 nM

In vitro

CHIR-98014 inhibits human GSK-3β with K_i of 0.87 nM. CHIR-98014 is very effective in preventing murine and rat GSK-3. Although CHIR-98014 acts as a simple competitive inhibitor of ATP binding, it displays from 500-fold to >1000-fold selectivity for GSK-3 versus 20 other protein kinases including Cdc2, ERK2, Tie-2 and KDR. CHIR-98014 prevents Cdc2 with IC50 of 3.7 μM. However, CHIR 98014 reveals similar ptoency against the highly homologous ɑ and β isoforms of GSK-3, it is noteworthy that it stronly discriminated between GSK-3 and its closest homologs CDC2 and ERK2. Exposure of insulin receptor-expressing CHO-IR cells or primary rat hepatocytes to increasing concentrations of inhibitor CHIR98014 results in a two- to three-fold stimulation of the GS activity ratio above basal. The concentrations of CHIR-98014 giving rise to half-maximal GS stimulation (EC50) is 106 nM and 107 nM for CHO-IR and rat hepatocytes, respectively. ^[1]

In vivo

GSK-3 inhibitor CHIR-98014 activates the GS activity ratio in isolated type I skeletal muscle from insulin-sensitive lean Zucker and from insulin-ressitant ZDF rats. Soleus muscle isolated from ZDF rats shows significant resistance to insulin for activation of GS but responded to 500 nM CHIR-98014 to the same extent (40% increase) as muscle from lean Zucker rats. Notably, GS activation by insulin plus CHIR-98014 is additive in muscle from lean Zucker rats and greater than additive in muscle from the ZDF rats. Total GS activity is not altered by either CHIR-98014 or insulin in these cells and muscles. Meanwhile, CHIR-98014 does not influence the insulin dose-response in muscle from lean animals. The reduction in hyperglycemia and improved glucose disposal are not limited to db/db mice and ZDF rats, as similar results are observed with ob/ob mice, diet-induced diabetic C57BL/6 mice, and glucose-intolerant SHHF rats treated with CHIR-98014. ^[1] Additionally, CHIR-98014 decreases the phosphorylation (Ser396) of tau protein in the cortex and hippocampus of postnatal rats. ^[2]

Protocol (from reference)

Kinase Assay:^{^[1]}	Kinase assays Polypropylene 96-well plates are ﬁlled with 300 μL/well buffer (50 mM tris HCl, 10 mM MgCl₂, 1 mM EGTA, 1 mM dithiothreitol, 25 mM β-glycerophosphate, 1 mM NaF, 0.01% BSA, pH 7.5) containing kinase, peptide substrate, and any activators. CHIR-98014 or controls are added in 3.5 μL of DMSO, followed by 50 μL of ATP stock to yield a ﬁnal concentration of 1 μM ATP in all cell-free assays. After incubation, triplicate 100-μL aliquots are transferred to Combiplate eight plates containing 100 μL/well 50 μM ATP and 20 mM EDTA. After 1 hour, the wells are rinsed ﬁve times with PBS, ﬁlled with 200 μL of scintillation ﬂuid, sealed, left 30 min, and counted in a scintillation counter. All steps are performed at room temperature.
Cell Assay:^{^[1]}	Cell lines CHO-IR cells Concentrations 0.01-10 μM Incubation Time 30 min Method CHO-IR cells expressing human insulin receptor are grown to 80% conﬂuence in Hamm’s F12 medium with 10% fetal bovine serum and without hypoxanthine. Trypsinized cells are seeded in 6-well plates at 1 × 10⁶ cells/well in 2 mL of medium without fetal bovine serum. After 24 hours, medium is replaced with 1 mL of serum-free medium containing GSK-3 inhibitor CHIR 98014 or control (ﬁnal DMSO concentration 0.1%) for 30 min at 37 °C. Cells are lysed by freeze/thaw in 50 mM tris (pH 7.8) containing 1 mM EDTA, 1 mM DTT, 100 mM NaF, 1 mM phenylmethylsulfonyl ﬂuoride, and 25 μg/mL leupeptin (buffer A) and centrifuged 15 min at 4 °C/14000 g. The activity ratio of GS is calculated as the GS activity in the absence of glucose-6-phosphate divided by the activity in the presence of 5 mM glucose-6-phosphate.
Animal Study:^{^[1]}	Animal Models db/db mice with 8-9 weeks Dosages 30 mg/kg Administration Subcutaneously

References

Customer Product Validation

: Data from [Stem Cells Dev, 2013, 22, 1893-90]

: Data from [Stem Cells Dev, 2013, 22, 1893-90]

: Data from [Stem Cells Dev, 2013, 22, 1893-90]

: , , Cancer Res, 2013, 73(4):1340-51.

Selleck's CHIR-98014 has been cited by 50 publications

RNA m6A modification regulates L1 retrotransposons in human spermatogonial stem cell differentiation in vitro and in vivo [ Cell Mol Life Sci, 2024, 81(1):92]	PubMed: 38363375
PIM1 drives lipid droplet accumulation to promote proliferation and survival in prostate cancer [ Oncogene, 2024, 43(6):406-419]	PubMed: 38097734
Inhibition of DNMT3B expression in activated hepatic stellate cells overcomes chemoresistance in the tumor microenvironment of hepatocellular carcinoma [ Sci Rep, 2024, 14(1):115]	PubMed: 38168140
CDKAL1 Drives the Maintenance of Cancer Stem-Like Cells by Assembling the eIF4F Translation Initiation Complex [ Adv Sci (Weinh), 2023, 10(12):e2206542]	PubMed: 36786012
mTORC1 Mediates Biphasic Mechano-Response to Orchestrate Adhesion-Dependent Cell Growth and Anoikis Resistance [ Adv Sci (Weinh), 2023, e2307206.]	PubMed: 38041494
Symmetric control of sister chromatid cohesion establishment [ Nucleic Acids Research,, 2023, 51(10)]	PubMed: None
Symmetric control of sister chromatid cohesion establishment [ Nucleic Acids Res, 2023, gkad146]	PubMed: 36912084
Irreversible cell cycle exit associated with senescence is mediated by constitutive MYC degradation [ Cell Rep, 2023, 42(9):113079]	PubMed: 37656618
Irreversible cell cycle exit associated with senescence is mediated by constitutive MYC degradation [ Cell Rep, 2023, 42(9):113079]	PubMed: 37656618
iPSC-derived exosomes promote angiogenesis in naturally aged mice [ Aging (Albany NY), 2023, 15(12):5854-5872]	PubMed: 37367945

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SHIPPING AND STORAGE
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