CHIR-99021 (CT99021) HCl

For research use only.

Catalog No.S2924

279 publications

CHIR-99021 (CT99021) HCl Chemical Structure

CAS No. 1797989-42-4

CHIR-99021 (CT99021) HCl is hydrochloride of CHIR-99021, which is a GSK-3α/β inhibitor with IC50 of 10 nM/6.7 nM; CHIR-99021 shows greater than 500-fold selectivity for GSK-3 versus its closest homologs Cdc2 and ERK2. CHIR-99021 is a potent pharmacological activators of the Wnt/beta-catenin signaling pathway. CHIR-99021 significantly rescues light-induced autophagy and augments GR, RORα and autophagy-related proteins.

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Selleck's CHIR-99021 (CT99021) HCl has been cited by 279 publications

Purity & Quality Control

Choose Selective GSK-3 Inhibitors

Biological Activity

Description CHIR-99021 (CT99021) HCl is hydrochloride of CHIR-99021, which is a GSK-3α/β inhibitor with IC50 of 10 nM/6.7 nM; CHIR-99021 shows greater than 500-fold selectivity for GSK-3 versus its closest homologs Cdc2 and ERK2. CHIR-99021 is a potent pharmacological activators of the Wnt/beta-catenin signaling pathway. CHIR-99021 significantly rescues light-induced autophagy and augments GR, RORα and autophagy-related proteins.
Targets
GSK-3β [1]
(Cell-free assay)
GSK-3α [1]
(Cell-free assay)
6.7 nM 10 nM
In vitro

CHIR-99021 shows greater than 500-fold selectivity for GSK-3 versus its closest homologs CDC2 and ERK2, as well as other protein kinases. Furthermore, CHIR-99021 shows only weak binding to a panel of 22 pharmacologically relevant receptors and little inhibitory activity against a panel of 23 nonkinase enzymes. CHIR-99021 induces the activation of glycogen synthase (GS) in insulin receptor-expressing CHO-IR cells with EC50 of 0.763 μM. [1] In addition to simulating the actions of insulin, inhibition of GSK-3 by CHIR-99021 (3 μM) increases free cytosolic β-catenin by 1.9-fold, mimicking the canonical Wnt signaling pathway in 3T3-L1 preadipocytes. During any of the first 3 days of differentiation, CHIR-99021 treatment inhibits the preadipocyte differentiation with IC50 of 0.3 μM by blocking induction of CCAAT/enhancer-binding protein α (C/EBPα) and peroxisome proliferator-activated receptor γ (PPARγ). [2] Unlike lithium chloride and AR-A014418, CHIR-99021 treatment does not reduce the viability of INS-1E cells even at high concentrations. Instead, CHIR-99021 robustly increases the rate of proliferation of INS-1E cells in a dose-dependent manner, and significantly inhibits INS-E cell death induced by high glucose and high palmitate in a concentration-dependent manner. CHIR-99021 promotes primary beta cell replication in isolated rat islets at concentrations as low as 1 μM, with 2-3 fold increase of cell replication at 5 μM of CHIR-99021 treatment. [3]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
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HEC-1 M4W1XGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NEPQOJpKSzVyPUGyPU4yQThizszN NH:wcW89[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmw1OTJzNEKvK|5USU6JRWK8M4E,
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HAL-01 MmXwS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NESw[FNKSzVyPUGyNk41QTlizszN MlyxQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xd4f3MoVjcS6jYz71b{9kcGWvYnyvZ49ueG:3bnTfdoVxd3K2X3PhdoQwS0iHTVLMOFEzOTR{Lze+V2FPT0WUPD;hQi=>
KYSE-140 MofqS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NHHuU5JKSzVyPUGyNk4yPDFizszN M4LqSlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4f3e{5m[mlwYXOueYsw[2inbXLsM4NwdXCxdX7kY5JmeG:{dG;jZZJlN0OKRV3CUFQyOjF2Mj:nQnNCVkeHUkyvZV4>
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NCI-N87 NWfyT3B6T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= Mnm1TWM2OD1zMkGuNFQ1KM7:TR?= MXi8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQly0NVIyPDJxJ{7TRW5ITVJ:L3G+
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769-P MYHHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NX7WeFhzUUN3ME2wMlU5PDl7IN88US=> NXvDXndQRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNPDF{MUSyM{c,W0GQR1XSQE9iRg>?
GCT NIHKb3BIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MV7JR|UxRTBwM{C2O|Ih|ryP NGK2Zmw9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmw1OTJzNEKvK|5USU6JRWK8M4E,
SW982 M1HhNWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NUm0NIx6UUN3ME2wMlIyPDF4IN88US=> NX3ZWJJZRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:ve5d4NmWkaT7hZ{52cy:laHXtZoww[2:vcH;1coRgemWyb4L0Y4NiemRxQ1jFUWJNPDF{MUSyM{c,W0GQR1XSQE9iRg>?
ES-CCE NU\RbVdLTnWwY4Tpc44hSXO|YYm= MXi1xsDPxE1? Ml7LN{Bl MXrEUXNQ NXzxSmxiW3S{b37ncJkhcW6mdXPld{B1cGViYXP0bZZqfHlib3[gW451N2KndHGtZ4F1\W6rbjDwZZRpf2G7 MXi8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDd5OUO2OUc,OjR5N{mzOlU9N2F-
ES-D3 MorhSpVv[3Srb36gRZN{[Xl? MnTJOeKh|ryP MnTWN{Bl M{e1VWROW09? NGjjenRUfHKxbnfsfUBqdmS3Y3XzJJRp\SCjY4Tpeol1gSCxZjDXcpQw[mW2YT3jZZRmdmmwIIDheIh4[Xl? MYK8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDd5OUO2OUc,OjR5N{mzOlU9N2F-
ES-CCE M1jHZ2N6fG:2b4jpZ{BCe3OjeR?= NGDKU4EyOMLizszN M2LoWVMh\A>? MkX2SG1UVw>? NYDORY5ZUW6mdXPld{Bud2SncnH0[UBkgXSxdH;4bYNqfHl? MWS8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDd5OUO2OUc,OjR5N{mzOlU9N2F-
ES-D3 M{PWOWN6fG:2b4jpZ{BCe3OjeR?= MkjvNVDDqM7:TR?= MkPFN{Bl MULEUXNQ MXrJcoR2[2W|IH3v[IVz[XSnIHP5eI91d3irY3n0fUB4cXSqIFnDOVAhd2ZiND65JO69VQ>? M2W1d|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2N{e5N|Y2Lz5{NEe3PVM3PTxxYU6=
Neural progenitor cells NH\tOXhHfW6ldHnvckBCe3OjeR?= NXP3Wo1EOyEQvF2= MV2xNEBl MX\EUXNQ M324c2lv\HWlZYOg[4Vv\XKjdHnvckBw\iCwZYXyZYwheHKxZ3XubZRweiClZXzsdy=> NV\lfYF7RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS2N|gxOzRpPkK0OlM5ODN2PD;hQi=>
A549 NYi5OZZUSXCxcITvd4l{KEG|c3H5 NYnyeFJRPSEEtV2= NVPXNWJJOjRiaB?= M{nPSGROW09? MYrJcoNz\WG|ZYOgZ4VtdCCjcH;weI9{cXN? MXK8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDN4N{[4PEc,OjR|Nke2PFg9N2F-
H157 NHrP[WRCeG:ydH;zbZMhSXO|YYm= NGfDRpo2KML3TR?= NYnjV|dyOjRiaB?= MVfEUXNQ NGnMN5hFd2W|IH7veEB{cWewaX\pZ4FvfGy7IHHm[oVkfCCjcH;weI9{cXN? NYLtfGtIRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkSzOlc3QDhpPkK0N|Y4Pjh6PD;hQi=>
H460 NUO0cVZJSXCxcITvd4l{KEG|c3H5 MnHGOUDDvU1? NGDYOFMzPCCq MX;EUXNQ MnzMRZR1\W63YYTld{Bk\WyuIHHwc5B1d3Orcx?= NXPaeG8{RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkSzOlc3QDhpPkK0N|Y4Pjh6PD;hQi=>
H157 MXfHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NHniW5g2KML3TR?= NUf2fHRzOjRiaB?= MlfXSG1UVw>? MlqyVJJwdW:2ZYOgZ4VtdCCycn;sbYZmemG2aX;u NVfLSotCRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkSzOlc3QDhpPkK0N|Y4Pjh6PD;hQi=>
H460 MXrHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MmnNOUDDvU1? MnjhNlQhcA>? MnS1SG1UVw>? M4DGPHBzd22xdHXzJINmdGxicILvcIln\XKjdHnvci=> Mof1QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR|Nke2PFgoRjJ2M{[3Olg5RC:jPh?=
embryonic stem cells (ESC) MYjGeY5kfGmxbjDBd5NigQ>? M2fEdFMhyrWP NXmyVpNmOTFiZB?= MkfaSG1UVw>? NF;1TYxRem:vb4Tld{B{\WyoLYLlcoV4[Wxib3[gcoHEt3[nLYT5dIUhdW:3c3Wg[Y1jenmxbnnjJJN1\W1iY3XscJMhMEWVQzm= MnPGQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjN6NEOzNVIoRjJ|OESzN|EzRC:jPh?=
DN3 MoHzSpVv[3Srb36gRZN{[Xl? MYKxNEDPxE1? NEHhcok6PiCq MlrRSG1UVw>? NXOxU3pOUW6lcnXhd4V{KGOnbHyg[IV3\WyxcH3lcpQh[W6mIIDyc4xq\mW{YYTpc44> M4PrbFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ|NUK2PVg6Lz5{M{WyOlk5QTxxYU6=
platelets MWDGeY5kfGmxbjDBd5NigQ>? Mn[yNVAh|ryP NHvOUJcyPSCvaX6= NHziW2JFVVOR NX3hcJRqUW6lcnXhd4V{KHSqcn;tZolvNW2nZHnheIVlKHCuYYTlcIV1KGGpZ4Ll[4F1cW:wIHHu[EBCXFBic3XjdoV1cW:w M3rrdlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ|MkO5PFc4Lz5{M{KzPVg4PzxxYU6=
hiPSC-NPs NYnONI1RT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= M4\kW|PjiIoQvF2= NWjmT3JlPzJiaB?= MlvZSG1UVw>? M4nQT2lv[3KnYYPld{Bk\WyuIIDyc4xq\mW{YYTpc47DqA>? MmSxQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjJ4NEK2PFcoRjJ{NkSyOlg4RC:jPh?=
INS-1E MWDGeY5kfGmxbjDBd5NigQ>? Mn7wNk42KM7:TR?= NEHqN3czPCCq MX3EUXNQ NIT5RWNKdmS3Y3XzJIEhemWmdXP0bY9vKG:oIF7QNUBwfmW{ZYjwdoV{e2mxbh?= MWC8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOjR{N{ewOEc,OjJ2Mke3NFQ9N2F-
PANC1 MYDBdI9xfG:|aYOgRZN{[Xl? NVLXWpdPyqB3IN88US=> M4PuZVczKGh? NYqzRmVPTE2VTx?= NIXpTFdVemmpZ3Xyd{BidiCjcH;weI91cWNicnXzdI9ve2V? NFS3XpA9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MkKwNVE5Pid-MkKyNFEyQDZ:L3G+
Endothelial cell(EC) MUjGeY5kfGmxbjDBd5NigQ>? MYGzJO69VQ>? MUDEUXNQ MWjQdo9ud3SnczDlcoRwfGinbHnhcEBk\WyuIDjFR{kh\GmoZnXy[Y51cWG2aX;u MUS8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOjB3MkCwOkc,OjJyNUKwNFY9N2F-
HMC1.2 MlX4S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? MmfLN|Ah|ryP MU[3NkBp NH\GPW5FVVOR MoDNVoVlfWOnczDj[YxtKHO3co\peoFt M1fjWlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ{MEO5N|AyLz5{MkCzPVMxOTxxYU6=
Sca-1+ HPC MlLqS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NWrhTJozOiEQvF2= M33FdlYh\A>? M4m1UGROW09? M1;YdHN6dmW{Z3nzeIlk[WyueTDwdo9ud3SnczDTZ4EuOSwEoFjQR{Bxem:uaX\ldoF1cW:wIHHu[EBkd2yxbomg[o9zdWG2aX;uJJRw\2W2aHXyJJdqfGhiRVfG NUjuXIp3RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkG2NlQ{PDhpPkKxOlI1OzR6PD;hQi=>
Oct4/Sox2/Klf4-infected MEFs MVTGeY5kfGmxbjDBd5NigQ>? MUCxNEDPxE1? M3\FTFI5KGR? NVjSXWg6TE2VTx?= MWPQdo9ud3SnczDNSWZ{KHKncILv[5JidW2rbnegWJJidnOmdXPl[EBjgSCRY4S0MEBUd3h{LDDhcoQhU2yoNB?= M4nIcVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF7OEO5NFU2Lz5zOUizPVA2PTxxYU6=
H4IIE MoXGSpVv[3Srb36gRZN{[Xl? M3HEd|ExKM7:TR?= MXOyOEBp MYjEUXNQ NIjsTItT\We3bHH0[ZMh|rJvY3H0[Y5qdiCjY4Tpeol1gSCjbnSgWGlTTSClb370ZYlvcW6pIIDyc41wfGW{IHHjeIl3cXS7 M1;rNVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF3M{WwNVk2Lz5zNUO1NFE6PTxxYU6=
insulin receptor–expressing CHO-IR NGPVcnVHfW6ldHnvckBCe3OjeR?= MoG3OVAxKG6P M4HXc|MxKG2rbh?= MkPBSG1UVw>? MXPBZ5RqfmG2ZYOgS4x6[2:pZX6gV5lvfGijc3WgLGdUMQ>? NGTyV4c9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zMk[wOlQ6Pyd-MUK2NFY1QTd:L3G+
3T3-L1 NXHCSHhTTnWwY4Tpc44hSXO|YYm= MUezJO69VQ>? MVu0JIg> NXLRb5U4TE2VTx?= NYHzVYJUUW6qaXLpeJMheHKnYXTpdI9kgXSnIHTp[oZmemWwdHnheIlwdg>? NG\jfWg9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zMkC1OVIxOCd-MUKwOVUzODB:L3G+
CHO-IR NXjye|VTTnWwY4Tpc44hSXO|YYm= NUL0cVBHOzBibXnu NXf2V4J6TUN3ME2wMlc4KM7:TR?= MYC8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zQTBzNkGyNUc,OjlyMU[xNlE9N2F-
HepG2 NY\QSFBvTnWwY4Tpc44hSXO|YYm= M4e2b|MhcA>? NXX2VpE1TUN3ME2xMlUh|ryP MXe8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOjJ4MUCyN{c,OjJ{NkGwNlM9N2F-

... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Western blot
Cleaved PARP / CD31 / VE-Cadherin / α-SMA / vimentin; 

PubMed: 30709379     


NCI-H460 cells were co-cultured with HUVECs in a 3D culture system, and then treated with 1 μM CHIR-99021, 10 μM or 20 μM Cisplatin, or a combination of both, for 24 h. Cells were harvested and immunoblotted with anti-cleaved PARP, anti-CD31, anti-VE-cadherin, anti-α-SMA, anti-vimentin, and anti-β-actin antibodies. 

CTNNB1; 

PubMed: 23993102     


Treatment with the GSK3β inhibitor CHIR-99021 led to increased levels of β-catenin in RKO and HT29 (non-mutant) cells.

β-catenin; 

PubMed: 24165128     


HeLa cells were treated with 5 μm CHIR-99021 for the indicated times. The samples were analyzed by Western blot.

30709379 23993102 24165128
Growth inhibition assay
Cell viability; 

PubMed: 22039301     


HMC-1.2 cells were cultured in the absence or presence of GSK3β inhibitor (CHIR 99021, 3–30 µM) and viability cell count was assessed by trypan blue dye exclusion after 24 h, 48 h or 72 h. All data are presented as the mean ± SEM of three independent experiments conducted in triplicate. p < 0.05*, p < 0.001** p < 0.0001*** vs vehicle treated control cells.

22039301
Immunofluorescence
SOX10 / EdU; 

PubMed: 28174705     


Incorporation of EdU (white arrowheads) into SOX10+ ENS (enteric nervous system) cells with and without CHIR-99021.

Oct3/4; 

PubMed: 24779365     


Fluorescence micrographs showing the protein expression of Brachyury/Oct3/4 after a three day differentiation of ES-D3 cells with BIO (0.5 μM), SB-216763 (5 μM), CHIR-99021 (5 μM), or CHIR-98014 (1 μM). Nuclei were counterstained with DAPI (blue). The merge is shown in yellow. Original magnification 400x. Scale bar = 50 μm.

28174705 24779365
In vivo Oral administration of CHIR-99021 at 30 mg/kg enhances glucose metabolism in a rodent model of type 2 diabetes, with a maximal plasma glucose reduction of nearly 150 mg/dl 3-4 hours after administration, while plasma insulin remains at or below control levels. Oral administration of CHIR-99021 at 16 or 48 mg/kg 1 hour before oral glucose challenges in ZDF rats significantly improves glucose tolerance with 14% and 33% reduction in plasma glucose at 16 mg/kg and 48 mg/kg, respectively, and the higher dose of CHIR-99021 also reduces hyperglycemia before the oral glucose challenge. [1] Administration of CHIR-99021 significantly augments hematopoietic repopulation in recipient mice transplanted with mouse or human hematopoietic stem cells (HSCs), suggesting that GSK-3 is a specific modulator of HSC activity. [4]

Protocol

Kinase Assay:

[1]

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Cell-free 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 27 nM GSK-3α or 29 nM GSK-3β, and 0.5 μM biotin-CREB peptide substrate. Various concentrations of CHIR-99021 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 minutes, and counted in a scintillation counter. All steps are performed at room temperature.
Cell Research:

[3]

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  • Cell lines: INS-1E
  • Concentrations: Dissolved in DMSO, final concentrations ~ 20 μM
  • Incubation Time: 1, or 4 days
  • Method:

    Cells are maintained for 24 hours in starvation medium (culture medium with only 5 mM glucose, 1% fetal calf serum). Then cells are exposed to various concentrations of CHIR-99021 for 1, or 4 days. Cell number is measured by staining of cellular DNA with CyQuant dye, which becomes fluorescent when bound to DNA. Fluorescence is measured after 30 minutes of incubation using the FLUOstar Optima reader. Cell replication is determined by BrdUrd incorporation. BrdUrd labeling solution is added to the medium for the last 4 hours before the cells are fixed using FixDenat solution and incubated with monoclonal anti-BrdUrd-POD antibodies. After substrate solution is added to each well, the light emission is measured in a microplate luminometer using the Analyst HT detection system.


    (Only for Reference)
Animal Research:

[1]

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  • Animal Models: Female db/db mice or male ZDF rats with type 2 diabetes
  • Dosages: ~48 mg/kg
  • Administration: Orally
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 93 mg/mL (185.33 mM)
Ethanol 2 mg/mL (3.98 mM)
Water Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
5% DMSO+30% PEG 300+ddH2O
For best results, use promptly after mixing.
24mg/mL

* 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.

Chemical Information

Molecular Weight 501.8
Formula

C22H18Cl2N8.HCl

CAS No. 1797989-42-4
Storage powder
in solvent
Synonyms N/A
Smiles CC1=CN=C(N1)C2=CN=C(N=C2C3=C(C=C(C=C3)Cl)Cl)NCCNC4=NC=C(C=C4)C#N.Cl

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
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Calculate the mass, volume or concentration required for a solution. The Selleck molarity calculator is based on the following equation:

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*When preparing stock solutions, please always use the batch-specific molecular weight of the product found on the via label and MSDS / COA (available on product pages).

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Calculate the dilution required to prepare a stock solution. The Selleck dilution calculator is based on the following equation:

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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

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Frequently Asked Questions

  • Question 1:

    I recently purchased CHIR 99021 to inject mice intraperitoneally. The data sheet says using 10% captisol for in vivo use. Is it for IP injection? Or do you have other solutions to suggest to use for in vivo IP injection of CHIR 99021?

  • Answer:

    The formulation of 15% Captisol is for oral gavage, and S2924 CHIR-99021 (CT99021) HCl in it is a suspension. S2924 CHIR-99021 (CT99021) HCl can be dissolved in 5% DMSO+30% PEG 300+ddH2O at 24mg/ml as a clear solution for IP injection.

  • Question 2:

    How about the half-life of this compound? If it is necessary for daily treatment when the compound is used in long-term experiment?

  • Answer:

    Please change the culture medium each day to add the new comound in it if the duration is more than 72h.

  • Question 3:

    What is the difference between S2924 and S1263? Which one is better for differentiation?

  • Answer:

    S1263 is the free base format of Chir-99021, while S2924 is the HCl salt format. They have the same biological activity but are slightly different in solubility.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID