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( f ) protein expression in lysates from 3D AKP organoids treated 5FU for 72 h, representative of n = 3 biological replicates
Biotinylated goat anti-mouse IgG or goat anti-rabbit IgG (Vector Labs, BA-9200 1:200 and BA-1000 1:200, respectively) secondary antibodies were added in blocking buffer and incubated for 1 h at room temperature
The following considerations further strengthen the criticism toward the use of PAC as a safer alternative to NSAID: PAC has been preferred to NSAIDs and steroids for the symptomatic and domiciliary management of the early stages of COVID-19, a choice grounded of the precautionary principle (but still theoretical in this specific case) principle primum non nocere (Micallef et al., 2020a)
MorrisGWalkerAJWalderKBerkMMarxWCarvalhoAFet al
PVDF membranes underwent blocking in 5% non-fat milk solution prior to overnight incubation with primary antibodies at 4 C
Approximately 150 g of digested peptide from each sample was used for purification, which is within the C18 spin columns maximum binding capacity of 100 g (#744101, Harvard Apparatus, Holliston, MA, USA) and then lyophilized before being analyzed on the Orbitrap Fusion Tribrid mass spectrometer coupled with an Ultimate 3000 nano-UPLC system (Thermo Fisher Scientific, Waltham, MA, USA)
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