34
YOSHINORI OTSUKI AND YUKO ITO
- wire loop
- injector with a filter (0.2 /-lm in pore size)
- Petri dishes
- Parafilm® (American National Can TM, Connecticut, U.S.A.)
- filter paper
Reagents - Fixative
- O.lM Phosphate buffer (PB) (pH7.4)
- 2% paraformaldehyde
- 2.5% glutaraldehyde
- Post-fixative
- 1% osmium tetroxide in 0.1 M PB
- Epoxy resin (Total volume 20 ml)
Epon 812
MNA
DDSA
DMP-30
9.41 m!
5.65 m!
4.94 m!
0.4 m!
- saturated sodium metaperiodate: 19 sodium metaperiodate
in 8ml distilled water
- Equilibration Buffer for the ISEL technique
- 30 mM Tris-HCI (pH 7.2)
- 140 mM sodium cacodylate
- 1 mM cobalt chloride
- 0.3 UI /-ll TdT for the ISEL technique
- Reaction Buffer for the ISEL technique: 0.04 nmol digoxigenin-dUTP, dATP
- Stop/Wash Buffer for the ISEL technique
- 300 mM sodium chloride
- 30 mM sodium citrate
- Tris-BSA buffer
- 0.02 M Tris-HCI buffer (pH7.4)
- 0.1 % Bovine Serum Albumin
- 10% normal sheep serum in Tris-BSA
- 10 nm colloidal gold conjugated to sheep anti-digoxigenin
antibody (e.g., British BioCell Int, Golden Gate, UK)