TdT 1 microliters (negative control does not include TdT)
Biotin 1 microliter
Total volume: 99 ul. This is enough for several coverslips or tissue section.
Remove slides from incubator and wash off TUNEL mixture thoroughly with 1X PBS
Put on streptavidin secondary (488 or Cy3) for 30 minutes room temperature.
Wash 3 times with 1X PBS
At this point you can label with additional antibodies. Minimally, samples should be counterstained with Hoechst or other nuclear dye. Proceed as directed for cell or tissue labeling. Otherwise coverslip with Gelvatol.
Specific staining should be nuclear and co-localize with your nuclear counterstain. Non-specific staining will be cytoplasmic. Avoid imaging the edges of samples