Dear everyone, I have paraffin embedded bone tissue (demineralized) and I would like to stain it with DAPI. What is the most simple protocol? I found many protocols online with antigen retrieval steps, blocking steps... I am guessing all I need is the permealization step with triton x-100 0.2 % in PBS for example. Do I also need a blocking step? if so, for what? All I need is the DAPI staining, I do not need any other antibodies.
Please someone direct me to a simple protocol, I would be very grateful.
Thank you so much
All the best
xylene (3 min) twice, 100 % ethanol (1:1) 3min, rehydration (ethanol 100 (3min), ethanol 95 (3min) ethanol 70 (3 min), ethanol 50 (3 min) ethanol 30 (3 min); permeablize ( triton x-100 0.2 % in PBS) 7 min; wash in PBS (5 min); wash in PBS (5 min); DAPI (1min-2min); wash in PBS (5 min) twice; dehydration with ethanol, xylen and mounting.
Thank you Nour Yahfoufi ! I will try.
Does the DAPI need to be added in the dark?
Yes in the dark you can cover any box with aluminium foil that should work
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