A. Remove 100µl from Tube 2 (Label solution) for 2 negative controls (50µl each). Do this even if you are omitting this negative control so that volumes and concentrations will remain consistent for the labeling.
B. Add Total volume (50µl,) of Tube 1 (TdT) + remainder of Tube 2 (450µl).
5ml 100mM Tris, pH 8.2
1 drop Levamisole
2 drops each of Solution 1, 2, and 3 of either Vector substrate
10mM Tris-HCl
20 mg/ml Proteinase K
100µl/slide x-phosphate/BCIP or Fast Red substrate
DNase I solution (1mg/ml - 1µg/ml) for Positive control
100mM Tris-Hcl, pH 8.2
4% Paraformaldehyde/PBS, pH 7.4
0.1% Triton X-100 in 0.1% Sodium Citrate (SSC)
2ml 20x SSC
400µl Triton X-100
398ml H2O
100mM Tris-Hcl, pH 8.2