Section8:Appendix 1. Immunoblot vs. Cytoblot Assays for Enzyme Inhibitor Potencies

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Enzyme Inhibition: Western Analysis

Enzyme Inhibition: C-Elisa Analysis

Enzyme Inhibitor Profiling

1. Immunoblot analysis. Confluent HEK293 cells were serum-starved for 2h before incubating with increasing concentrations of specific enzyme inhibitors 1, 2 &3 for 1h. Where indicated, PMA was added for 10 min. Cell lysates were collected, electrophoresed and blotted. Blots were stained with a degenerate antibody to the specific enzyme phosphorylation consensus sequence that recognizes the phosphorylated form of several substrates in these cells.
2. C-ELISA was performed on parallel cultures using duplicate wells per data point.