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Cells were incubated with the agonist for 15 min (except in the case of KRAS assays, where the incubation time was 60 min), and bioluminescence resonance energy transfer (BRET) measurements were taken using a Victor X4 plate reader (PerkinElmer) immediately after the addition of coelenterazine h (final concentration: 1 M)
To ale neznamen, e ho mme vdy dostatek
This represents a precautionary approach rather than a requirement based on proven deficiency risk
The dose stays the same at 1000 mcg (1 mg) per injection
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