Hello,
I am working with an Exiqon miCURY LNA microRNA ISH kit tot stain some types of miRNA in cardiac mouse tissue sections (parrafine).
The ISH procedure works splendid! The probes are DIG labeled and when I amplify it immunohistochemically with αDIG-AP and then use NBT-BCIP I see perfect staining in my positive controls and no staining at al in my negative controls. Exactly how it should look like and really clear.
When I try to amplify this ISH fluorescent it goes wrong... I cant get any signal at all.
I use a PerkinElmer TSA PLUS Fluorescence kit tot amplify my signal. I have used FITS labeld TSA and Cy5 labaled TSA, both give me no result on the ISH. But the TSA works perfect! When I use TSA in respons on another staining with HRP conjugated secondary Ab's, this give me perfect results as well.
So you would expect there is something wrong with my αDIG-HRP Ab's (Roche).... but they are brand new and work fine in other reactions.
Where can it go wrong?
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