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CHIR-99021 (CT99021)

Catalog No.
A3011
GSK-3 inhibitor, Cell-permeable, ATP-competitive
Grouped product items
SizePriceStock Qty
10mM (in 1mL DMSO)
$83.00
In stock
Evaluation Sample
$30.00
In stock
5mg
$77.00
In stock
10mg
$116.00
In stock
25mg
$231.00
In stock
100mg
$611.00
In stock
For scientific research use only and should not be used for diagnostic or medical purposes.

Tel: +1-832-696-8203

Email: [email protected]

Worldwide Distributors

Background

CHIR-99021 is a highly specific glycogen synthase kinase-3 (GSK-3) inhibitor which can inhibit both isoforms with IC50 of 10 nM (GSK-3α) and 6.7 nM (GSK-3β).

CHIR-99021 was proved to promote self-renewal and maintain pluripotency of both B6 and BALB/c ES cells via stabilizing the downstream effectors like c-Myc and β -catenin[1]. In J1 mESC cells, CHIR-99021 played an important role in maintaining the colony morphology as well as the self-renewal when combined with leukemia inhibitory factor (LIF). CHIR-99021 has shown to regulate multiple signaling pathways which involve Wnt/β-catenin, TGF-β, Nodal and MAPK, and the expression of epigenetic regulatory genes like Dnmt3l[2].

CHIR-99021 has been demonstrated to promote DN3 thymocytes proliferation and differentiation in the absence of pre-TCR signaling, Notch1 signaling or CXCL12[3]. However, study has also found that higher concentration (10 ?M but not 1 ?M or 3 ?M) of CHIR99021 might selectively inhibit differentiation by activating IL-7 signaling pathway[3].

References:
1.?Ye S1, Tan L, Yang R, Fang B, Qu S, Schulze EN, Song H, Ying Q, Li P. Pleiotropy of glycogen synthase kinase-3 inhibition by CHIR99021 promotes self-renewal of embryonic stem cells from refractory mouse strains. PLoS One. 2012;7(4):e35892.
2.?Wu Y1, Ai Z, Yao K, Cao L, Du J, Shi X, Guo Z, Zhang Y. CHIR99021 promotes self-renewal of mouse embryonic stem cells by modulation of protein-encoding gene and long intergenic non-coding RNA expression. Exp Cell Res. 2013
3.?PLoS One. 2013;8(3):e58501. doi: 10.1371/journal.pone.0058501. Epub 2013 Mar 20. Schroeder JH1, Bell LS, Janas ML, Turner M. Pharmacological inhibition of glycogen synthase kinase 3 regulates T cell development in vitro. PLoS One. 2013;8(3):e58501.

Product Citation

Chemical Properties

Physical AppearanceA solid
StorageStore at -20°C
M.Wt465.34
Cas No.252917-06-9
FormulaC22H18Cl2N8
SynonymsCHIR99021, CHIR-99021, CHIR 99021, CT99021,GSK-3 Inhibitor XVI
Solubility≥23.27 mg/mL in DMSO; insoluble in H2O; insoluble in EtOH
Chemical Name6-((2-((4-(2,4-dichlorophenyl)-5-(5-methyl-1H-imidazol-2-yl)pyrimidin-2-yl)amino)ethyl)amino)nicotinonitrile
SDFDownload SDF
Canonical SMILESN#CC1=CC=C(NCCNC2=NC=C(C3=NC=C(C)N3)C(C4=CC=C(Cl)C=C4Cl)=N2)N=C1
Shipping ConditionSmall Molecules with Blue Ice, Modified Nucleotides with Dry Ice.
General tips We do not recommend long-term storage for the solution, please use it up soon.

Protocol

Cell experiment: [1]

Cell lines

Human embryonic stem cells (ESCs)

Preparation method

The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37 °C for 10 minutes and/or shake it in the ultrasonic bath for a while.Stock solution can be stored below -20°C for several months.

Reaction Conditions

8 μM, 24 hours

Applications

On day 0, differentiation was initiated with 8 μM CHIR-99021 for 24 h conferring canonical Wnt/β-catenin activation, followed by Wnt inhibition on day 3 by addition of 4 μM IWR-1 for 48 h. For all groups tested, first signs of eGFP-fluorescence and first beating EBs were observed on day 6 with a constant increase until d10, reaching almost 100% of beating EBs for the groups seeded with 666-2000 cells per aggregate. Flowcytometry analysis of dissociated EBs on day 10 showed the highest yield of 5.9 Nkx2-5-eGFP+ cells for the group seeded with 666 cells/aggregate; higher cell numbers per aggregate resulted in lower yields. Immunofluorescence stainings of EB cryosections and dissociated/reseeded cells confirmed a high content of Nkx2-5+ and cTnT+ cardiomyocytes, thereby demonstrating efficient cardiomyogenic differentiation of human ESC-derived EBs after aggregation on agarose microwells and induction with small molecule-based media.
Animal experiment: [2]

Animal models

Akita type 1 diabetic mice and wild-type mice

Dosage form

Intraperitoneal injection, 50 mg/kg, daily

Applications

At 15 min after the propranolol injection, the 2-min average HF fraction increased from 46.8 ± 2.9% before CHIR-99021 treatment to 67.8 ± 5.1% after CHIR-99021 treatment. Treatment of Akita mice with CHIR-99021 increased SREBP-1 from 0.53 ± 0.07- to 1.17 ± 0.11- fold. CHIR-99021 treatment increased GIRK4 levels from 0.28 ± 0.06- to 1.08 ± 0.14- fold of those in WT mice, which was significantly higher than in placebo.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

[1] Dahlmann J, Kensah G, Kempf H, et al. The use of agarose microwells for scalable embryoid body formation and cardiac differentiation of human and murine pluripotent stem cells. Biomaterials, 2013, 34(10): 2463-2471.

[2] Zhang Y, Welzig C M, Picard K L, et al. Glycogen synthase kinase-3β inhibition ameliorates cardiac parasympathetic dysfunction in type 1 diabetic Akita mice. Diabetes, 2014, 63(6): 2097-2113.

Biological Activity

Description CHIR-99021 (CT99021) is an inhibitor of GSK-3α/β with IC50 of 10 nM/6.7 nM; > 500-fold selectivity for GSK-3 versus its closest homologs CDC2 and ERK2, as well as other protein kinases.
Targets GSK-3          
IC50 7 nM          

Quality Control

Chemical structure

GSK-3 Inhibitor XVI(CHIR 99021)

Related Biological Data

CHIR-99021
Intracellular?antibody FACS staining for Nestin and TUJ1 indicated a stark contrast in the number of neurons generated by LSB (2% TUJ1+) compared to LSB3i (75% TUJ1+). When one or two of the three inhibitors used in 3i were added, the same level of TUJ1 cells was not achieved; however, CHIR (CHIR99021) with either SU5402 or DAPT achieved >53% neurons, indicating a requirement for CHIR in the formation of TUJ1+ neurons. Results are presented as mean ± s.e.m.

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