[Histonet] Melanin Bleach

[Maxim Peshkov]

Karen,
The bleaching reagents will not compatible before IHC anyway.
Some tips:
1. Do IHC-test as usual and counterstain nuclei with methylene blue Loeffler at 3-5 sec, only for
profile of chromatine. The melanin will stain at green color.
2. Use any red color chromogen for IHC. It will be some contrast with brown melanine
3. Cut sections at 3 microns. Do all steps of IHC as usual, include DAB.
All next steps do very gently:
Before nuclear counterstain do bleaching procedure with 0.01% KMnO4 + 0.5% H2SO4 at 10 mins;
gently rinse in DW 1 min;
0.5% oxalic acid 5-10 sec or up to bleaching;
gently rinse in DW 1 min;
counterstain nuclei and all other steps with regular manner of your lab.
This solutions will not bleach DAB.
Sections will not detach from slides.

Hope this help.
-- 
 Maxim Peshkov
 Russia
 Taganrog.                         mailto:Maxim_71 at mail.ru