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Recovery: Recovery rates for amino acids ranged from 80.68% to 103.54%, and for acylcarnitines from 93.37% to 108.35%, confirming acceptable performance

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After incubation, and washes with PBS, appropriate secondary fluorescent antibodies (1:500, AlexaFluor-conjugated, Invitrogen) were added to the blocking solution at room temperature for 2 h in the dark

Accordingly, we next employed preclinical models of kidney injury to gain mechanistic insights into the potential interplay between TMAO and kidney function and the putative molecular nature of the 2nd hit