Catalog No.S7198 Synonyms: GSK-3 Inhibitor IX, 6-bromoindirubin-3-oxime

BIO Chemical Structure

Molecular Weight(MW): 356.17

BIO is a specific inhibitor of GSK-3 with IC50 of 5 nM for GSK-3α/β in a cell-free assay, shows >16-fold selectivity over CDK5, also a pan-JAK inhibitor.

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  • Lysates of HCT116p53KO cells were harvested 24 hs after treatment with different GSK3 inhibitors and GSK3A/B activation/inactivation checked by western blot: a mix of pSer21-GSK3A and pSer9-GSK3B antibodies and antibody cross-reacting with both pTyr279-GSK3A and pTyr216-GSK3B were used to assess the specificity of the inhibitor for GSK3A. BIO: 6-bromoindirubin-3'-oxime, TWS: TWS119, SB2: SB216763, SB4: SB415286.

    PLoS One 2014 9(7), e100947. BIO purchased from Selleck.

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Biological Activity

Description BIO is a specific inhibitor of GSK-3 with IC50 of 5 nM for GSK-3α/β in a cell-free assay, shows >16-fold selectivity over CDK5, also a pan-JAK inhibitor.
Features The first pharmacological agent shown to maintain self-renewal in human and mouse embryonic stem cells.
GSK-3 [1]
(Cell-free assay)
TYK2 [4]
(Cell-free assay)
CDK5/p35 [1]
(Cell-free assay)
CDK2/CyclinA [1]
(Cell-free assay)
CDK1/CyclinB [1]
(Cell-free assay)
5 nM 30 nM 0.08 μM 0.30 μM 0.32 μM
In vitro

BIO (6-bromoindirubin-3'-oxime) is a specific inhibitor of glycogen synthase kinase-3 (GSK-3), with IC50 of 5 nM for GSK-3α/β, shows >16-fold selectivity over CDK5. BIO interacts within the ATP binding pocket of these kinases, reduces β-catenin phosphorylation on a GSK-3-specific site in cellular models, closely mimicks Wnt signaling in Xenopus embryos. [1] In human and mouse embryonic stem cells, BIO maintains the undifferentiated phenotype and sustains expression of the pluripotent state-specific transcription factors Oct-3/4, Rex-1 and Nanog. BIO-mediated Wnt activation is functionally reversible, as withdrawal of the compound leads to normal multidifferentiation programs in both human and mouse embryonic stem cells. [2] BIO promotes proliferation in mammalian cardiomyocytes. [3]6BIO is also a pan-JAK inhibitor, with IC50 values of 0.03, 1.5, 8.0, 0.5 μM for TYK2, JAK1, JAK2 and JAK3. BIO selectively inhibits phosphorylation of STAT3 and induces apoptosis of human melanoma cells. [4]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
human SH-SY5Y cells MXjGeY5kfGmxbjDhd5NigQ>? NXjrS2ZTUW6qaXLpeIlwdiCxZjDHV2s{NW2nZHnheIVlKGKndHGtZ4F{\WmwIIDoc5NxcG:{eXzheIlwdiCrbjDoeY1idiCVSD3TXVV[KGOnbHzzJIlvKHC{ZYPlcoNmKG:oIF3HNVMzKGK7IGfld5Rmem5iYnzveEBidmGueYPpd{whUUN3ME2wMlI6KM7:TR?= M4\Ee|E5QDF4MUGw
human K562 cells NUnhT246WHKxbHnm[ZJifGmxbjDhd5NigQ>? NWjV[IdtPzJiaB?= MVvBcpRqeHKxbHnm[ZJifGm4ZTDhZ5Rqfmm2eTDh[4FqdnO2IHj1cYFvKEt3NkKgZ4VtdHNiYX\0[ZIhPzJiaILzJIJ6KE2WVDDhd5NigSxiSVO1NF0yNjNizszN NGnTdm8yQTd6M{G0PS=>
human IMR90 cells NXe5[oJLWHKxbHnm[ZJifGmxbjDhd5NigQ>? MmrHO|IhcA>? Mn;6RY51cXC{b3zp[oVz[XSrdnWgZYN1cX[rdImgZYdicW6|dDDoeY1idiCLTWK5NEBk\WyuczDh[pRmeiB5MjDodpMh[nliTWTUJIF{e2G7LDDJR|UxRTFwOTFOwG0> NG\lTIQyQTd6M{G0PS=>
human HCT116 cells M1TNWXBzd2yrZnXyZZRqd25iYYPzZZk> M1rTPVczKGh? NG\ScmdCdnSrcILvcIln\XKjdHn2[UBi[3Srdnn0fUBi\2GrboP0JIh2dWGwIFjDWFEyPiClZXzsd{Bi\nSncjC3NkBpenNiYomgUXRVKGG|c3H5MEBKSzVyPUWuNkDPxE1? NXnyOIdYOTl5OEOxOFk>
human HL60 cells MmC1VJJwdGmoZYLheIlwdiCjc4PhfS=> M1OzVVUh\GG7cx?= NYHuNHc5SW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[WejaX7zeEBpfW2jbjDIUFYxKGOnbHzzJIFnfGW{IEWg[IF6eyCkeTDNWHQh[XO|YYm= NV\TOpFMOTl5OEOxOFk>
human HuH7 cells MmTNVJJwdGmoZYLheIlwdiCjc4PhfS=> NHfWTmQ4OiCq MUXBcpRqeHKxbHnm[ZJifGm4ZTDhZ5Rqfmm2eTDh[4FqdnO2IHj1cYFvKEi3SEegZ4VtdHNiYX\0[ZIhPzJiaILzJIJ6KE2WVDDhd5NigQ>? NWXsWlduOTl5OEOxOFk>
human SH-SY5Y cells NXjKNnVmS3m2b4TvfIlkyqCjc4PhfS=> NH;H[|E1QCCq M{[2fGN6fG:2b4jpZ4l1gSCjZ3HpcpN1KGi3bXHuJHNJNVO\NWmgZ4VtdHNiYX\0[ZIhPDhiaILzJIJ6KE2WUzDy[YR2[3Srb36gZZN{[XluIFnDOVA:QSEQvF2= M3jwXFE5QDF4MUGw
human NB39 cells MWXDfZRwfG:6aXRCpIF{e2G7 NV3HbVdIPDhiaB?= MkXCR5l1d3SxeHnjbZR6KGGpYXnud5QhcHWvYX6gUmI{QSClZXzsd{Bie3Onc4Pl[EBieyClZXzsJJZq[WKrbHn0fUBi\nSncjC0PEBpenNiYomgUXRVKGG|c3H5 M{T0c|IyQDB{OUS3
human SK-N-SH cells MoPER5l1d3SxeHnjxsBie3OjeR?= NEftd4Y1QCCq NHrsVFhEgXSxdH;4bYNqfHliYXfhbY5{fCCqdX3hckBUUy2QLWPIJINmdGy|IHHzd4V{e2WmIHHzJINmdGxidnnhZoltcXS7IHHmeIVzKDR6IHjyd{BjgSCPVGSgZZN{[Xl? MXuyNVgxOjl2Nx?=
mouse HT22 cells M2rGVmZ2dmO2aX;uJIF{e2G7 NWDke4V7OTBizszN NXnmdVhwOjRiaB?= NYC4cZp5UW6qaXLpeIlwdiCxZjDHV2s{NW2nZHnheIVlKGKndHGgZ4F{\WmwIIDoc5NxcG:{eXzheIlwdiCrbjDtc5V{\SCKVEKyJINmdGy|IHH0JFExKHWPIHHmeIVzKDJ2IHjyd{BjgSCZZYP0[ZJvKGKub4SgZY5idHm|aYOgbY4heHKnc3XuZ4Uhd2ZiTVexN|I> MmHlNlI6QTh2NEO=
HEK293 cells NXrwbGVWTnWwY4Tpc44h[XO|YYm= NFjtOFExNjVvMTFOwG0> MkjOTY5pcWKrdHnvckBw\iCqdX3hckBIW0t|IHHjeIl3cXS7IHnuJGhGUzJ7MzDj[YxteyClb370ZYlvcW6pIITo[UBYdnRxYnX0ZU1k[XSnbnnuJIFkfGm4YYTl[EBz\XCxcoTldkBxW3WyZYLUU3BHVEGVSDCoV3RHOjl|IHPlcIx{MSCjdDCwMlUhfG9iMTD1UUBjgSCZboSgdoVxd3K2ZYKg[4Vv\SCjc4PhfS=> NG\Re4ozPDZ7N{K0OC=>

... Click to View More Cell Line Experimental Data

In vivo BIO suppresses melanoma tumor growth in a mouse xenograft model. [4]


Kinase Assay:[1]
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Kinase assay:

Kinase activities are assayed in Buffer A or C at 30°C, at a final ATP concentration of 15 μM. Blank values are subtracted and activities calculated as pmoles of phosphate incorporated during a 10 min incubation. Controls are performed with appropriate dilutions of dimethylsulfoxide. In a few cases phosphorylation of the substrate is assessed by autoradiography after SDS-PAGE. GSK-3α/β is purified from porcine brain by affinity chromatography on immobilized axin. It is assayed, following a 1/100 dilution in 1 mg BSA/ml 10 mM DTT, with 5 μl 40 μM GS-1 peptide, a specific GSK-3 substrate, (YRRAAVPPSPSLSRHSSPHQSpEDEEE), in buffer A, in the presence of 15 μM [γ-32P] ATP (3,000 Ci/mmol; 1 mCi/ml) in a final volume of 30 μl. After 30 min incubation at 30°C, 25 μl aliquots of supernatant are spotted onto 2.5 × 3 cm pieces of Whatman P81 phosphocellulose paper, and 20 seconds later, the filters are washed five times (for at least 5 min each time) in a solution of 10 ml phosphoric acid/liter of water. The wet filters are counted in the presence of 1 ml ACS scintillation fluid.
Cell Research:[1]
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  • Cell lines: COS1, Hepa or SH-SY5Y cells
  • Concentrations: ~10 μM
  • Incubation Time: 12 or 24 h
  • Method: COS1, Hepa (wild-type, CEM/LM AhR deficient and ELB1 ARNT deficient), or SH-SY5Y cells are grown in 6 cm culture dishes in Dulbecco's Modified Medium (DMEM) containing 10% fetal bovine serum. For treatment, IO (5 μM), BIO (5 or 10 μM), MeBIO (5 or 50 μM), LiCl (20 or 40 mM), or mock solution (DMSO, 0.5% final concentration) is added to medium when cell density reaches ∼70% confluence. After 12 (SH-SY5Y) or 24 hours, the cells, while still in plate, are lysed with lysis buffer (1% SDS, 1 mM sodium orthovanadate, 10 mM Tris [pH 7.4]). The lysate is passed several times through a 26G needle, centrifuged at 10,000 × g for 5 min, and adjusted to equal protein concentration. About 8 μg of each sample is loaded for immunoblotting. Enhanced chemiluminescence is used for detection. The following primary antibodies are used: mouse anti-β-catenin CT (Upstate Biotechnolgies, Clone 7D8, recognizes total β-catenin), mouse anti-phospho-β-catenin (Upstate Biotechnologies, Clone 8E7, recognizes dephosphorylated β-catenin), mouse anti-GSK-3 β, mouse anti-GSK-3 phosphoTyr216, rabbit anti-AhR (Aryl hydrocarbon receptor), and rabbit anti-actin.
    (Only for Reference)
Animal Research:[4]
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  • Animal Models: mouse
  • Formulation: freshly prepared in 30% Solutol (Basf) at a concentration of 10 mg/mL.
  • Dosages: 50 mg/kg
  • Administration: Oral gavage
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 71 mg/mL (199.34 mM)
Ethanol 6 mg/mL (16.84 mM)
Water Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
30% PEG400+0.5% Tween80+5% propylene glycol
For best results, use promptly after mixing.
30 mg/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 356.17


CAS No. 667463-62-9
Storage powder
in solvent
Synonyms GSK-3 Inhibitor IX, 6-bromoindirubin-3-oxime

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Clinical Trial Information

NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT03607409 Recruiting Infertility IVI Middle East Fertility Clinic September 9 2018 --
NCT03462108 Recruiting Safety Issues PT Bio Farma April 9 2018 Phase 1
NCT03022708 Recruiting Heart Defect Congenital Xeltis May 8 2017 Not Applicable
NCT03398148 Recruiting Ulcerative Colitis (UC) AbbVie March 7 2018 Phase 2|Phase 3
NCT03345849 Recruiting Crohn''s Disease AbbVie December 7 2017 Phase 3
NCT03337763 Recruiting Heart Failure Biotronik SE & Co. KG November 7 2017 --

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GSK-3 Signaling Pathway Map

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID