CK2 inhibitor (Ki = 0.25 μM), more selective than DMAT. Unlike DMAT, TMCB displays selectivity over PIM1, HIPK2 and DYRK1a (Ki values are 8.65, 15.25 and 11.90 μM, respectively) and a more favourable selectivity profile over a range of other kinases.
Store at Room Temperature. The product can be stored for up to 12 months.
Soluble in DMSO to 25 mM (with heating)
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.
ab23423 staining cIAP2 in HeLa cells treated withTMCB (ab120289), by ICC/IF. Decrease in cIAP2 expression correlates with increased concentration of TMCB, as described in literature. The cells were incubated at 37°C for 10 minutes in media containing different concentrations of ab120289 (TMCB) 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 ab23423 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat 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.
Functional Studies - TMCB (ab120289)
HeLa cells were incubated at 37°C for 24h with vehicle control (0 µM) and different concentrations of TMCB (ab120289). Decreased expression of cIAP2 in HeLa cells correlates with an increase TMCB concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 40 µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab23423 at 2 µg/ml and ab1801 at 2 µg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.