Does anyone have a protocol or any useful tips for inhibiting transcription in mammalian cell lines with Actinomycin D?

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Kostas Iatrou

Actinomycin-D (not B) will largely inhibit ribosomal RNA synthesis. For inhibition of RNA Pol II (mRNA synthesis) you should use alpha-amanitin.

Celina Lafaille

While it is true that Actinomycin-D will also inhibit RNA Pol I, it is also very efficient with Pol II, and its inhibition kinetics are way faster than alpha-amanitin, so I would stick with Actinomycin-D rather than alpha-amanitin