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The cells were moistened twice with seahorse-specific test medium and incubated in a non-CO 2 incubator for 1 h

Volume_30ml

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Because ESI often adds one or more protons (H⁺, mass ~1.008), the instrument will detect ions like [M+H]⁺ at m/z ≈ 1420.5, [M+2H]²⁺ at m/z ≈ 710.75, or [M+3H]³⁺ at m/z ≈ 474.1