CHIR 99021

Name: CHIR 99021
Brand: Tocris Bioscience
SKU: 4423
Rating: 5 (16 reviews)

Tocris Bioscience, part of Bio-Techne | Catalog # 4423

(16)

Citations (1456)

COA / SDS

Highly selective GSK-3 inhibitor; acts as Wnt activator

Key Product Details

Description:	Highly selective GSK-3 inhibitor; acts as Wnt activator
Alternative Names:	CHIR99021,CT99021
Chemical Name:	6-[[2-[[4-(2,4-Dichlorophenyl)-5-(5-methyl-1H-imidazol-2-yl)-2-pyrimidinyl]amino]ethyl]amino]-3-pyridinecarbonitrile
Purity:	≥98% (HPLC)
Molecular Weight:	465.34

View Full Technical Details for CHIR 99021

Technical Data
Citations / Reviews (16)

Biological Activity for CHIR 99021

CHIR 99021 is a potent and highly selective inhibitor of glycogen synthase kinase 3 (GSK-3) (IC₅₀ values are 6.7 and 10 nM for GSK-3β and GSK-3α respectively). Acts as Wnt activator. Exhibits >500-fold selectivity for GSK-3 over closely related kinases; also displays >800-fold selectivity against 45 additional enzymes and receptors. In combination with the irreversible inhibitor of LSD1 (Cat. No. 3852), CHIR 99021 enables reprogramming of mouse embryonic fibroblasts, transduced by Oct4 and Klf4 only, into iPSCs. Enhances mouse and human ESC self-renewal when used in combination with PD 0325901 (Cat. No. 4192). It promotes differentiation of hPSCs in cardiomyocytes and allows formation of extended pluripotent stem (EPS) cells in combination with (S)-(+)-Dimethindene maleate (Cat. No. 1425), tetracycline antibiotic (Cat. No. 3268) and human leukemia inhibitory factor. Used in a protocol to generate brain organoids from human iPSCs. Also used in iPS cell differentiation protocol that produces CD34⁺ hematopoietic cells with multilineage engraftment ability.

CHIR 99021 synthesized to cGMP guidelines and CHIR 99021 in solution also available.

For more information on how to use CHIR 99021, see our protocols: Transdifferentiating Fibroblasts into Neurons, Culturing Transcriptomically Distinct Pluripotent mESCs, Converting Fibroblasts into Cardiomyocytes (9C Cocktail), Human iPSC-derived Myoblasts.

Compound Libraries for CHIR 99021

CHIR 99021 is also offered as part of the Tocriscreen 2.0 Max, Tocriscreen Kinase Inhibitor Library, Tocriscreen Epigenetics Library and Tocriscreen Stem Cell Library. Find out more about compound libraries available from Tocris.

External Portal Information

Chemicalprobes.org is a portal that offers independent guidance on the selection and/or application of small molecules for research. The use of CHIR-99021 is reviewed on the chemical probes website.

Scientific Data Examples for CHIR 99021

	Cardiomyocyte derived from human induced pluripotent stem cells (hiPSCs) using CHIR 99021. Cells cultured in Media supplemented with Activin A (Catalog # 338-AC, R&D Systems), N21-MAX Insulin Free (Catalog # AR010, R&D Systems), BMP-4 (Catalog # 314-BP, R&D Systems), bFGF (Catalog # 233-FB, R&D Systems) and DKK-1 (Catalog # 5439-DK, R&D Systems). Stained for F-Actin (Phalloidin, green), Troponin T (red, Human Troponin T (Cardiac) Antibody, Catalog # MAB1874, R&D Systems) and counterstained with DAPI (blue, Catalog # 5748, Tocris).

Technical Data for CHIR 99021

M. Wt	465.34
Formula	C₂₂H₁₈Cl₂N₈
Storage	Store at -20°C
Purity	≥98% (HPLC)
CAS Number	252917-06-9
PubChem ID	9956119
InChi Key	AQGNHMOJWBZFQQ-UHFFFAOYSA-N
Smiles	ClC(C=C1Cl)=CC=C1C2=NC(NCCNC3=CC=C(C#N)C=N3)=NC=C2C4=NC=C(C)N4

The technical data provided above is for guidance only. For batch specific data refer to the Certificate of Analysis.

Tocris products are intended for laboratory research use only, unless stated otherwise.

Solubility for CHIR 99021

	Solvent	Max Conc. mg/mL	Max Conc. mM
Solubility
Solubility	DMSO	9.31	20

Preparing Stock Solutions for CHIR 99021

The following data is based on the product molecular weight 465.34.

Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.

Select a batch to recalculate based on the batch molecular weight:

Concentration / Solvent Volume / Mass	1 mg	5 mg	10 mg
0.2 mM	10.74 mL	53.72 mL	107.45 mL
1 mM	2.15 mL	10.74 mL	21.49 mL
2 mM	1.07 mL	5.37 mL	10.74 mL
10 mM	0.21 mL	1.07 mL	2.15 mL

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References for CHIR 99021

References are publications that support the biological activity of the product.

Hoang Generation of spatial-patterned early-developing cardiac organoids using human pluripotent stem cells. Nat.Protoc. 2018 PMID: 29543795
Lian Robust cardiomyocyte differentiation from human pluripotent stem cells via temporal modulation of canonical Wnt signaling. Proc.Natl.Acad.Sci.U.S.A. 2012 PMID: 22645348
Wimmer Human blood vessel organoids as a model of diabetic vasculopathy. Nature 2019 PMID: 30651639
Noor 3D Printing of Personalized Thick and Perfusable Cardiac Patches and Hearts. Adv.Sci.(Weinh) 2019 PMID: 31179230
Dye In vitro generation of human pluripotent stem cell derived lung organoids. Elife 2015 PMID: 25803487
Koehler 3D mouse embryonic stem cell culture for generating inner ear organoids. Nat.Protoc. 2014 PMID: 24784820
Bartfeld In vitro expansion of human gastric epithelial stem cells and their responses to bacterial infection. Gastroenterology 2015 PMID: 25307862
Ring Selective glycogen synthase kinase 3 inhibitors potentiate Ins activation of glucose transport and utilization in vitro and in vivo. Diabetes 2003 PMID: 12606497
Sato SnapShot: Growing Organoids from Stem Cells. Cell 2015 PMID: 26091044
Mahe Establishment of gastrointestinal epithelial organoids Curr.Protoc. Mouse Biol. 2014 PMID: 25105065
Yang Derivation of pluripotent stem cells with in vivo embryonic and extraembryonic potency. Cell. 2017 PMID: 28388409
Ye Pleiotropy of glycogen synthase kinase-3 inhibition by CHIR99021 promotes self-renewal of embyronic stem cells from refractory mouse strains. PLoS One 2012 PMID: 22540008
Li Generation of human-induced pluripotent stem cells in the absence of exogenous Sox2. Stem Cells 2009 PMID: 19839055
Schafer An in vivo neuroimmune organoid model to study human microglia phenotypes. Cell 2023 PMID: 37172564
Xia The generation of kidney organoids by differentiation of human pluripotent cells to ureteric bud progenitor-like cells. Nat. Protoc. 2014 PMID: 25340442
Ng Long-term engrafting multilineage hematopoietic cells differentiated from human induced pluripotent stem cells. Nat.Biotechnol. 2024 PMID: 39223325
Pan AKT kinase activity is required for Li to modulate mood-related behaviors in mice. Neuropsychopharmacology 2011 PMID: 21389981