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1.) For quantification of staining by Flow Cytometry, what should I use as an isotype control? 2.) I would like to analyze live cells that are undergoing apoptosis, can you please suggest a viability dye that can be used with this kit? 3.) In step 1 of the staining procedure, you recommend preparing a negative control using Z-VAD-FMK to inhibit caspase activation. How long should I incubate my cells with this inhibitor?
Asked on Apr 12 2011
Thank you for contacting us. 1.) You may use Caspase-8 negative cells (cells that were not stimulated to induce Caspase-8) for your negative control in Flow Cytometry. 2.) For a viability dye, you may try using PI but this will need to be optimized for your applications. 3.) The Z-VAD-FMK inhibitor may be incubated for 30-60 minutes or for the same amount of time that your test inhibitor is incubated with the cells. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Answered on Apr 12 2011