1. PI3K/Akt/mTOR
    Stem Cell/Wnt
    Autophagy
  2. GSK-3
    Autophagy

CHIR-99021 (Synonyms: CT99021)

Cat. No.: HY-10182 Purity: 99.92%
Data Sheet SDS Handling Instructions

CHIR-99021 is a GSK-3α/β inhibitor 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.

For research use only. We do not sell to patients.
CHIR-99021 Chemical Structure

CHIR-99021 Chemical Structure

CAS No. : 252917-06-9

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Free Sample (0.5-1 mg)   Apply now  
10 mM * 1 mL in DMSO $66 In-stock
5 mg $60 In-stock
10 mg $90 In-stock
50 mg $270 In-stock
100 mg $430 In-stock
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500 mg   Get quote  

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Customer Review

Other Forms of CHIR-99021:

    CHIR-99021 purchased from MCE. Usage Cited in: Toxicology. 2016 May 20;355-356:31-38.

    Western analysis of β-catenin protein level in zebrafish embryos exposed to EOM in the presence or absence of CH/CHIR (n = 3). Different letters indicate significant differences. Moreover, both CH and CHIR attenuate the EOM-induced changes in the protein level of b-catenin and in EROD activity.

    CHIR-99021 purchased from MCE. Usage Cited in: Nat Med. 2016 May;22(5):547-56.

    Immunofluorescent staining for α-actinin (ACTN2) and cardiac troponin T (TNNT2) to demonstrate sarcomeric organization in hiPSC–CMs derived from patients who do not experience Doxorubicin–induced cardiotoxicity (DOX) versus those who do experience Doxorubicin-induced cardiotoxicity (DOXTOX) after 24 h treatment with Doxorubicin. Scale bar, 20 µm.

    CHIR-99021 purchased from MCE. Usage Cited in: Cell Signal. 2016 Mar;28(3):148-56.

    Inhibition of PI3K p110δ signalling, either by PI3KCD-/- or using PI3K p110δ inhibitor PI-3065, significantly blocks LPS-induced GSK3α phosphorylation.
    • Biological Activity

    • Protocol

    • Technical Information

    • Purity & Documentation

    • References

    Description

    CHIR-99021 is a GSK-3α/β inhibitor 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.

    IC50 & Target

    IC50: 10 nM/6.7 nM (GSK-3α/β)[1]

    In Vitro

    CHIR 99021inhibits human GSK-3β with Ki values of 9.8 nM[1]. CHIR 99021 is a small organic molecule that inhibits GSK3α and GSK3β by competing for their ATP-binding sites.In vitro kinase assays reveal that CHIR 99021 specifically inhibits GSK3β (IC50=~5 nM) and GSK3α (IC50=~10 nM), with little effect on other kinases[2]. In the presence of CHIR-99021 the viability of the ES-D3 cells is reduced by 24.7% at 2.5 μM, 56.3% at 5 μM, 61.9% at 7.5 μM and 69.2% at 10 μM CHIR-99021 with an IC50 of 4.9 μM[3].

    In Vivo

    In ZDF rats, a single oral dose of CHIR 99021 (16 mg/kg or 48 mg/kg) rapidly lowers plasma glucose, with a maximal reduction of nearly 150 mg/dl 3-4 h after administration[1]. CHIR99021 (2 mg/kg) given once, 4 h before irradiation, significantly improves survival after 14.5 Gy abdominal irradiation (ABI). CHIR99021 treatment significantly blocks crypt apoptosis and accumulation of p-H2AX+ cells, and improves crypt regeneration and villus height. CHIR99021 treatment increases Lgr5+ cell survival by blocking apoptosis, and effectively prevents the reduction of Olfm4, Lgr5 and CD44 as early as 4 h[4].

    References
    Preparing Stock Solutions
    Concentration Volume Mass 1 mg 5 mg 10 mg
    1 mM 2.1490 mL 10.7448 mL 21.4897 mL
    5 mM 0.4298 mL 2.1490 mL 4.2979 mL
    10 mM 0.2149 mL 1.0745 mL 2.1490 mL
    Please refer to the solubility information to select the appropriate solvent.
    Kinase Assay
    [2]

    Kinases are purified from SF9 cells through use of their His or Glu tag. Glu-tagged proteins are purified, and His-tagged proteins are purified. Kinase assays are performed in 96-well plates with appropriate peptide substrates in a 300-μL reaction buffer (variations on 50 mM Tris-HCl, pH 7.5, 10 mM MgCl2, 1 mM EGTA, 1 mMdithiothreitol, 25 mMβ-glycerophosphate, 1 mM NaF, and 0.01% bovine serum albumin). Peptides has Km values from 1 to 100 μM. CHIR 99021 or CHIR GSKIA is added in 3.5 μL of Me2SO, followed by ATP to a final concentration of 1 μM. After incubation, triplicate 100-μL aliquots are transferred to Combiplate 8 plates containing 100 μL/well of 50 μM ATP and 20 mM EDTA. After 1 hour, the wells are rinsed five times with phosphate-buffered saline, filled with 200 μL of scintillation fluid, sealed, and counted in a scintillation counter 30 min later. All of the steps are at room temperature. The percentage of inhibition is calculated as 100×(inhibitor-no enzyme control)/(Me2SO control-no enzyme control)[2]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    Cell Assay
    [3]

    CHIR 99021 is dissolved in DMSO and stored, and then diluted with appropriate media before use[3].

    The viability of the mouse ES cells is determined after exposure to different concentrations of GSK3 inhibitors for three days using the MTT assay. The decrease of MTT activity is a reliable metabolism-based test for quantifying cell viability; this decrease correlates with the loss of cell viability. 2,000 cells are seeded overnight on gelatine-coated 96-well plates in LIF-containing ES cell medium. On the next day the medium is changed to medium devoid of LIF and with reduced serum and supplemented with 0.1-1 μM BIO, or 1-10 μM SB-216763, CHIR-99021 or CHIR-98014. Basal medium without GSK3 inhibitors or DMSO is used as control. All tested conditions are analyzed in triplicates[3]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    Animal Administration
    [1][4]

    CHIR 99021 is formulated as solutions in 20 mM citrate-buffered 15% Captisol or as fine suspensions in 0.5% carboxymethylcellulose (Rat)[1].
    CHIR 99021 is prepared in DMSO and diluted (Mice)[4].

    Rat[1]
    Primary hepatocytes from male Sprague Dawley rats that weighed <140 g are prepared and used 1-3 h after isolation. Aliquots of 1×106cells in 1 mL of DMEM/F12 medium plus 0.2% BSA and CHIR 99021(orally at 16 or 48 mg/kg) or controls are incubated in 12-well plates on a low-speed shaker for 30 min at 37°C in a CO2-enriched atmosphere, collected by centrifugation and lysed by freeze/thaw in buffer A plus 0.01% NP40; the GS assay is again performed.
    Mice[4]
    Mice 6-10 weeks old are used. The PUMA+/+ and PUMA-/- littermates on C57BL/6 background (F10) and Lgr5-EGFP (Lgr5-EGFP-IRES-creERT2) mice are subjected to whole body irradiation (TBI), or abdominal irradiation (ABI). Mice are injected intraperitoneally (i.p.) with 2 mg/kg of CHIR99021 4 h before radiation or 1 mg/kg of SB415286 28 h and 4 h before radiation. Mice are sacrificed to collect small intestines for histology analysis and western blotting. All mice are injected i.p. with 100 mg/kg of BrdU before sacrifice. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    References
    Molecular Weight

    465.34

    Formula

    C₂₂H₁₈Cl₂N₈

    CAS No.

    252917-06-9

    Storage
    Powder -20°C 3 years
      4°C 2 years
    In solvent -80°C 6 months
      -20°C 1 month
    Shipping

    Room temperature in continental US; may vary elsewhere

    Solvent & Solubility

    DMSO: ≥ 5.1 mg/mL

    * "<1 mg/mL" means slightly soluble or insoluble. "≥" means soluble, but saturation unknown.

    Purity: 99.92%

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    Inquiry Information

    Product Name:
    CHIR-99021
    Cat. No.:
    HY-10182
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