I am working on tracing with HRP & WGA-HRP
I used TMB reaction to visualize the HRP/WGA-HRP.
However the staining is a bit uneven, and the blue staining can not be washed down with 0.01 Ace buffer.
Do anyone have other good protocols for HRP visualization? or with IHC, anti-WGA, anti-HRP reactions?
My detailed protocol as below:
1. wash sections in pure water, 6 times * 5 min
2. A solution + B solution mixture, 20-25 'C, 20 min incubate with sections
3. every 100 ml mixture solution, add 30 ul 30% H2O2, mix well and put the sections back for another 20-25 min
5. wash with 0.01 M Ace buffer , 0-4 'C, 6 times * 5 min
6. mount on to gelatin-coated slides.
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