Biological descriptionHighly potent, selective, reversible and ATP-competitive inhibitor of GSK3α/β (IC50 values are 5 (GSK3α/β), 83 (CDK5/p25), 300 (CDK2/cyclin A) and 320 nM (CDK1/cyclin B)). Activates the Wnt signaling pathway in vitro maintaining self-renewal and pluripotency in ESC. Cell-permeable.
Storage instructionsStore at -20°C. Store at -20°C long term. It is important to note that this product is reported to be light sensitive. Store In the Dark. Store under desiccating conditions.
Solubility overviewSoluble in DMSO to 25 mM
Wherever possible, you should prepare and use solutions on the same day. However, if you need to make up stock solutions in advance, we recommend that you store the solution as aliquots in tightly sealed vials at -20°C. Generally, these will be useable for up to one month. Before use, and prior to opening the vial we recommend that you allow your product to equilibrate to room temperature for at least 1 hour.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
ab32572 staining ßcatenin in SW480 cells treated with BIO (ab120891), by ICC/IF. Increase of ßcatenin expression correlates with increased concentration of BIO, as described in literature. The cells were incubated at 37°C for 48h in media containing different concentrations of ab120891 (BIO) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32572 (1/200) dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Polychronopoulos P et al. Structural basis for the synthesis of indirubins as potent and selective inhibitors of glycogen synthase kinase-3 and cyclin-dependent kinases. J Med Chem47:935-46 (2004).
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