Hello,
Im asking to check if anyone has a protocol for preparing fluorescent tyramide for fluorescent in situ hybridization. The one Im using isnt working, so if anyone has one that works, can you please link me to it or share it with me?
doi: 10.1007/978-1-4939-3622-9_4. Hope this article will help you.
Hi Maroun Abi Younes,
I would recommend to use the fluorescent TSA reagents from Akoya Biosciences (previously from PerkinElmer), with 1X Plus Amplification Diluent. I don't know on which samples you're working, but it is suitable for FISH in free-floating mouse brain sections.
If you need to synthesize it yourself, this website describes how to prepare everything (Fluorophore-Tyramide and amplification diluent): https://sites.google.com/site/helobdellaprotocols/histology/tsa
If I remember, you also asked a question related to the dissolution of Tyramine. I did not have problem with that using the protocol described in the link above. Make sure to mix 10µL of Triethylamine (Sigma T0886) in 1mL of anhydrous N,N-dimethylformamide (Sigma 227056) and use this solution to dissolve the Tyramine-HCl (Sigma T2879) at 10mg/mL.
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