APOPTAG KIT REAGENTS
3% H2O2 in PBS
Proteinase K
10 ug/ml in PBS
30% H2O2 PBS Chick: st 12
st 14
st 16
st 24
none
3 min
5 min
15 min
5 ml
45 ml
10
90
20
180
30
270
Mouse: thymus
16.5 gut
10 min
15 min
Note: Keep Equilibration Buffer, Reaction Buffer and TdT on ice while in use
Working Strength TdT Enzyme
The TdT enzyme must be diluted with Reaction Buffer prior to use. Vortex the TdT
enzyme before using. To prepare, add in a fresh microcentrifuge tube:
77 ul Reaction buffer
33 ul TdT enzyme (vortex before use)
110 ul Total
The smallest amount of working solution to use is 55ul per slide. Slides with more tissue
may require >55ul, but it is important to be as conservative as possible. Keep on ice up
to six hours. Vortex before applying to slides.
Working strength StopWash Buffer
Prepare by adding 1 ml StopWash Buffer to 34 ml dH2O. This reagent may be prepared
and stored in a glass or plastic container at 4oC for up to 1 year. Use a fresh aliquot for
each run.
Counterstain for tissue with LacZ
1. Diluted hematoxylin (Anatech normal strength) 1:10 in
1% aqueous glacial acetic acid
2. Dip in H2O
3. Scott’s Tap Water
4. Running H2O
5. Rinse in dH2O
6. Run to Xylene and mount
30 sec
10 dips
5 minutes