I will be treating mice of different genetic backgrounds with carcinogen, fixing their lungs with formalin, storing in 70%EtOH and performing TUNEL assay to determine differing levels of apoptosis. Is there a positive control treatment that I can use on the mice in place of carcinogen that is sure to induce apoptosis detected by TUNEL?
After sample collection, be sure to run positive and negative control slides for TUNEL on lung tissue. For the positive, a brief treatment with DNase after permeabilization will fragment enough DNA, EVERY nucleus should be positive. If they are not, you are not getting adequate tissue permeabilization. Optimize this first.
Next, use healthy tissue as a control, perform TUNEL _exactly_ as you did for the DNase positive (except without the DNase, obviously). This should have a very low level of TUNEL positive cells (I expect
Monocrotaline i(Sigma) nduces apoptosis in pulmonary endothelium in vivo at 600mg/kg subcutaneously. See Aliotta et al. Cardiovasc Res 2013 attached.
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