Apoptosis - caspase-3 activity (Oct/19/2005 )
iam a PhD student , working on the apoptosis in the reference of host defence in the case of
tuberculosis. For caspase-3 assay we are using substrate cojugated with pNA and AMC(7-amono-4methylcumarine) any body can tell me whats should be different between two groups reading (Normal and Infected cells caspase-3 levels) uring taking absorbance reading for pNA and flourescence reading for AMC conjugated substrates.
Thanks a lot.
I'm also a phd student doing a similar project, im looking for apoptosis occuring during the infection of macrophages. In my case 24hrs post infection I get caspase 3 activation at about 10% greater than the control of uninfected cells. I use AMC-DEVD caspase 3 substrate What mycobacteria do you use? and what macrophage cell line? Apparently Keane et al. suggests that greater amouts of apoptosis occurs with less virulent strains of mycobacteria.