Benzidine-peroxidase staining for blood (Conlon lab)
Protocol submitted by VGP from Conlon lab protocols [1]
link to protocol document [2]
Benzidine Staining
(Hemoglobin histochemical stain)
1) Make up 4X Benzidine stock
- 100 mg Benzidine
- 3.125 ml HAc (Glacial)
- 3.125 ml H2O
- store @ 4°C in the DARK
2) Fix Embryos in 1X benzidine for 5-10 minutes
- 500ul Benzidine/1480ul H2O
3) Add 20ul 30% H2O2
4) Stain 2-15 minutes
5) Wash several times in 12% Glacial Acetic Acid
6) Fix 1.5 h in MEMFA
7) Change MEMFA 3X
8) Wash 3-4X EtOH
9) Store @ 4°C or 20°C in EtOH