CHIR-98014

Catalog No.S2745 Batch:S274501

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Technical Data

Formula

C20H17Cl2N9O2
Molecular Weight 486.31 CAS No. 252935-94-7
Solubility (25°C)* In vitro DMSO 32 mg/mL (65.8 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.
* <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 Ki of 0.87 nM. This compound is very effective in preventing murine and rat GSK-3. Although it 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. This inhibitor prevents Cdc2 with IC50 of 3.7 μM. However, it reveals similar potency against the highly homologous ɑ and β isoforms of GSK-3, it is noteworthy that it strongly 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 this chemical results in a two- to three-fold stimulation of the GS activity ratio above basal. The concentrations 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 of this compound to the same extent (40% increase) as muscle from lean Zucker rats. Notably, GS activation by insulin plus this chemical 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 this inhibitor or insulin in these cells and muscles. Meanwhile, this compound 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 this chemical. [1] Additionally, this compound 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

  • https://pubmed.ncbi.nlm.nih.gov/12606497/
  • https://pubmed.ncbi.nlm.nih.gov/17906685/

Customer Product Validation

<p>Differentiation of hESCs along the vascular. (B) FACS analysis showing the percentage of cells expressing CD34, PDGFRa, and KDR after 4 days of differentiation using our differentiation protocol in H9 hESC line. Similarly, an analysis was provided for the H1 hESC line using the same protocol but with CHIR98014 as the GSKi.</p>

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

<p> </p><div>T (Brachyury) and b-catenin expression in hESCs after GSKi treatment. (C) Immunofluorescence analysis of OCT4 and brachyury in hESCs after 24 h of GSKi treatment. Top row: H1-hESCs treated with basal media alone without any GSKi, middle row: H9 hESCs treated with CHIR99021, bottom row: H1-hESCs treated with 2 mM CHIR98014. (D) 20 · magnification of bcatenin expression in H1-hESCs after 24 h of GSKi treatment. Top row: hESCs treated with basal media alone without any</div><div>GSKi, middle row: hESCs treated with CHIR99021, bottom row: hESCs treated with CHIR98014. All scale bars represent 200 mm.</div>

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

<p>T (Brachyury) and b-catenin expression in hESCs after GSKi treatment. (B) Comparison of transcription profiles for T and CXCR4 in hESCs treated using CHIR99021 at 5mM and a different GSKi, CHIR98014 at 2mM. Although gene expression levels were different, the transcription profiles for both genes were approximately similar for both GSKis with T upregulating and peaking at day 1, while CXCR4 begins up-regulating at day 2.</p>

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

<p>Western blotting (D) in U937 cells following 16-hour exposure to BEZ235/ABT-737 (0.5 μmol/L each) in the presence or absence of BIO, MeBIO, or CHIR-98014 (2 μmol/L each).</p>

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

Selleck's CHIR-98014 Has Been Cited by 62 Publications

Sulforaphane-cysteine inhibits α-tubulin/PD-L1/PFKFB4 axis leading to apoptosis in human glioblastoma [ Med Oncol, 2025, 42(8):333] PubMed: 40659958
Pantethine ameliorates dilated cardiomyopathy features in PPCS deficiency disorder in patients and cell line models [ Commun Med (Lond), 2025, 5(1):323] PubMed: 40745475
Nociceptor-immune interactomes reveal insult-specific immune signatures of pain [ Nat Immunol, 2024, 25(7):1296-1305] PubMed: 38806708
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
Identification of novel neuroprotectants against vincristine-induced neurotoxicity in iPSC-derived neurons [ Cell Mol Life Sci, 2024, 81(1):315] PubMed: 39066803
PIM1 drives lipid droplet accumulation to promote proliferation and survival in prostate cancer [ Oncogene, 2024, 43(6):406-419] PubMed: 38097734
Enhancing Maturation and Translatability of Human Pluripotent Stem Cell-Derived Cardiomyocytes through a Novel Medium Containing Acetyl-CoA Carboxylase 2 Inhibitor [ Cells, 2024, 13(16)1339] PubMed: 39195229
Enhancing Maturation and Translatability of Human Pluripotent Stem Cell-Derived Cardiomyocytes through a Novel Medium Containing Acetyl-CoA Carboxylase 2 Inhibitor [ Cells, 2024, 13(16)1339] PubMed: 39195229
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
Characterization of the BH1406 non-small cell lung cancer (NSCLC) cell line carrying an activating SOS1 mutation [ Transl Lung Cancer Res, 2024, 13(11):2987-2997] PubMed: 39670010

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