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closePotential artifacts
Posted by tcvanaman on 02 Apr 2008 at 15:31 GMT
The CaM kinase inhibitor STO 609 is used at a concentration far too high to be specific for CaM Kinase Kinase. At 5 umolar, STO 609 would inhibit many kinases (see Sigma catalog). The concentration of testosterone used to initiate apoptosis is also far beyond physiologial concentrations (see JBC 2006 281:25492-25501.).
RE: Potential artifacts
cguest replied to tcvanaman on 02 Apr 2008 at 21:07 GMT
The STO-609 dose we chose to use for CaMKK inhibition was selected based on the work of Tokumitsu (J Biol Chem. 2002 May 3;277(18):15813-8 and J Biol Chem. 2003 Mar 28;278(13):10908-13). Tokumitsu showed that CaMKK was potently inhibited at 5 ug/ml (not umolar) without significantly altering PKC activity. Even at a concentration double the one we used, PKC activity was not significantly altered.
We did not conduct any experiments measuring apoptosis using testosterone.