CHIR 99021

Tocris Bioscience | Catalog # 4423

GMP Version Available: TB4423-GMP
GMP
Highly selective GSK-3 inhibitor; acts as Wnt activator
Tocris Bioscience

Key Product Details

Description

Highly selective GSK-3 inhibitor; acts as Wnt activator

Alternative Names

CHIR99021,CT99021
View all GSK-3 Inhibitors »

Product Description

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 (see protocols below). 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, Reprogramming MEFs into Skeletal Muscle Progenitors, highly-efficient-generation-cipscs-mefs, Generation of β cells from hPSCs, generation-and-propagation-eps-cells-lcdm-cocktail, generating-vascular-organoids, 3D Culture of Lung Alveolar Cells, Differentiation of Hematopoietic Progenitors from hPSCs, Generating Midbrain Dopaminergic Neurons from hPSCs, Generation of Kidney Organoids from hPSCs.

Scientific Data Images for CHIR 99021

Cardiomyocyte derived from human induced pluripotent stem cells (hiPSCs) using 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).

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.

Product Specifications for CHIR 99021

Molecular Weight

465.34

Formula

C22H18Cl2N8

Storage

Store at -20°C

Purity

≥98%

Chemical Name

6-[[2-[[4-(2,4-Dichlorophenyl)-5-(5-methyl-1H-imidazol-2-yl)-2-pyrimidinyl]amino]ethyl]amino]-3-pyridinecarbonitrile

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.

Solubility

Solvent Max Conc. mg/mL Max Conc. mM
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 all 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

Calculators

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Background References

References are publications that support the biological activity of the product. See our Citations tab to view 2030 publications citing the usage of this product.

Product Documents for CHIR 99021

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for CHIR 99021

For research use only

Related Research Areas

Notch Pathway

Customer Reviews for CHIR 99021 (19)

4.9 out of 5
19 Customer Ratings
5 Stars
89%
4 Stars
11%
3 Stars
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2 Stars
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1 Stars
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Customer Images

Showing  1 - 5 of 19 reviews Showing All
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  • Best for 2i LIF media - mESCs
    Name: Rahul Roy
    Species: Mouse
    Assay Type: In Vitro
    Verified Customer | Posted 05/08/2025
    Used for mESCs 2i LIF media
    CHIR 99021 4423
  • IPS culture
    Name: Murali K Yanda
    Species: Mouse
    Assay Type: In Vitro
    Cell Line/Tissue: ips
    Verified Customer | Posted 09/27/2022
    Mouse ips cells culture with 2i media.
    CHIR 99021 4423
  • 2i medium culture
    Name: Yu Chen
    Species: Mouse
    Assay Type: In Vitro
    Cell Line/Tissue: ESC
    Verified Customer | Posted 07/11/2022
    ESC 2i medium culture; 3 uM
    CHIR 99021 4423
  • ESC culture
    Name: Xiaoyu Zhang
    Species: Mouse
    Assay Type: In Vitro
    Cell Line/Tissue: ESC
    Verified Customer | Posted 11/23/2021
    In vitro cell culture addition. The image shows the morphology of ESC colonies after conversion using the treatment condition. This condition supported the conversion and long-term maintenance of dome-shaped ESCs from primed ESCs.
    CHIR 99021 4423
  • iPSC differentiation
    Name: Ricardo Malvicini
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: Human iPSC
    Verified Customer | Posted 09/26/2021
    Used for the differentiation of Human IPSc to cardiomyocites (concentrarion of 5uM)
  • Differentiation to cartilage
    Name: Julieta O'Flaherty
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: iPSCs
    Verified Customer | Posted 12/08/2020
    I used at 3uM to generate cartilage from iPSCs, worked really well. Keep at -80C for long term storage
    CHIR 99021 4423
  • Kidney organoids
    Name: Kirsty Rooney
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: MAN13 hESCs
    Verified Customer | Posted 11/21/2020
    I use 8uM of CHIR to differentiate hESCs to intermediate mesoderm and then to kidney organoids
    CHIR 99021 4423
  • Differentiation of human pluripotent stem cells
    Name: Paul Humphreys
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: hPSCs
    Verified Customer | Posted 11/20/2020
    Perform titration against your cell line to optimise correct concentration.
    Directed differentiation of human pluripotent stem cells (hPSCs) towards mesoderm lineages. Used at concentration of 3uM. Very effective in induction of mesoderm markers.
    CHIR 99021 4423
  • iPSC differentiation
    Name: Stephanie Hsiao
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: iPSC
    Verified Customer | Posted 10/13/2020
    iPSC culture medium supplemented with 4 uM of CHIR99021.
    Differentiation of human iPSCs into hematopoietic progenitor cells.
    CHIR 99021 4423
  • It worked good
    Name: Anonymous
    Species: Mouse
    Assay Type: In Vitro
    Cell Line/Tissue: iPSC
    Verified Customer | Posted 07/04/2020
    The cells were then maintained in RM 15 days with fresh media every other day
    Reprogramming media comprises of 3 μM CHIR99021 and 0.5 μM A-83-01.
    CHIR 99021 4423
  • For maintenance of stem cells pluripotency
    Name: Anonymous
    Species: Mouse
    Assay Type: In Vitro
    Cell Line/Tissue: Neural Stem Cells
    Verified Customer | Posted 06/25/2020
    It's better to aliquot and keep it at -80°C for long term storage.
    For directed differentiation to neural, cells are treated with GSK-3 inhibitor, CHIR99021 to induce canonical WNT signaling. This product gave me reproducible results.
    CHIR 99021 4423
  • Works as intended.
    Name: Anonymous
    Species: Rat
    Assay Type: In Vitro
    Cell Line/Tissue: Sprague-Dawley rats primary neuronal
    Verified Customer | Posted 04/04/2020
    10 μM for 2 h
    CHIR 99021 4423
  • Differentiation of human dopaminergic neurons from stem cells
    Name: Anonymous
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: Stem Cells
    Verified Customer | Posted 02/17/2020
    Used this product routinely to differentiate dopaminergic neurons from human stem cells.
    CHIR 99021 4423
  • The hiPSCs were maintained with Essential 8 Flex Medium with CHIR 99021
    Name: Anonymous
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: ipsc
    Verified Customer | Posted 02/08/2020
    1.4 μM
    CHIR 99021 4423
  • All iPSC lines and R1 mouse ES cells were maintained on feeders in KO-DMEM (Invitrogen) with 3 mM CHIR99021, and 1 mM PD0325901.
    Name: Anonymous
    Species: Mouse
    Assay Type: In Vitro
    Cell Line/Tissue: ipsc
    Verified Customer | Posted 12/04/2019
    All iPSC lines and R1 mouse ES cells were maintained on feeders in KO-DMEM (Invitrogen) with 3 mM CHIR99021, and 1 mM PD0325901.
    CHIR 99021 4423
  • works great for Wnt activation
    Name: avinash waghray
    Species: Human
    Assay Type: In Vitro
    Verified Customer | Posted 08/08/2019
    In Tissue culture for growing basal cells
  • Check the maintenance of naive pluripotency in 2i
    Name: Megha PB
    Species: Mouse
    Assay Type: In Vitro
    Cell Line/Tissue: E14TG2a
    Verified Customer | Posted 07/18/2018
    Culture mouse ES cells in 2i media and without 2i to assess differentiation. Only the cells in 2i are pluripotent while mES cells cultured in -2i for 48 hours exit naive pluripotency.
    CHIR 99021 4423
  • reliable product
    Name: Pinyuan Tian
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: pluripotent stem cells
    Verified Customer | Posted 07/05/2018
    Although the datasheet says store at -20°C, it's better to aliquot and keep it at -80°C for long term storage.
    For directed differentiation of pluripotent stem cells to kidney organoids, cells are treated with GSK-3 inhibitor, CHIR99021 to induce canonical WNT signaling. CHIR99021 concentration and differentiation time period are the key factors for this protocol. I found this product gave me reproducible results.
    CHIR 99021 4423
  • cells treatment
    Name: Anonymous
    Assay Type: In Vitro
    Cell Line/Tissue: Neuroblastoma
    Verified Customer | Posted 12/04/2017
    Treating cells

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