Hi guys,
I want to inhibit PKC epsilon with an inhibitor peptide (EAVSLKPT from Santa Cruz). Unfortunately, I found no relevant information about the appropriate permeabilisation protocol : some authors use saponin (at different concentrations) or triton etc...
Does anyone already used this inhibitor peptide ? What protocol did you use ? How long is the efficiency of the inhibition by the peptide ?
Thanks a lot for your help !
I am trying to solve the same problem. I need a PKCepsilon inhibitor that works in vivo.
You may want to use the cell-permeable inhibitor PKCe141 http://www.emdmillipore.com/US/en/product/PKC-Translocation-Inhibitor-II-PKCe141-Calbiochem,EMD_BIO-530993#overview
Also, the peptide can be coupled to TAT to make it cell permeable, but I haven't found a source yet. Amadesi S, Cottrell GS, Divino L, Chapman K, Grady EF, Bautista F, Karanjia R, Barajas-Lopez C, Vanner S, Vergnolle N, Bunnett NW (2006) Protease-activated receptor 2 sensitizes TRPV1 by protein kinase Cepsilon- and A-dependent mechanisms in rats and mice. J Physiol 575:555-571.
Hi Claudie, I also plan to use the PKC epsilon inhibitor peptide and was wondering if you found a solution for your problem?
Hi Markus,
Actually I tried to permeabilize my cells with saponin and it gave awful results... so I am now testing with digitonin for a very short time (1-5 min), and it seems really less toxic !
I am currently testing the efficacy of this permeabilization protocol with the peptide, I'll let you know.
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