Fig. 2: Design of fluorogenic probe for extracellular metabolite detection via NAD(P)H-coupled assay. | Communications Chemistry

Fig. 2: Design of fluorogenic probe for extracellular metabolite detection via NAD(P)H-coupled assay.

From: Development of pathway-oriented screening to identify compounds to control 2-methylglyoxal metabolism in tumor cells

Fig. 2

a Structure of Q-dsAMC and the design of live cell-based coupled assay. b Fluorescence spectra (λex. = 360 nm) of 1 μM probes (Q-dsAMC and dsAMC) in PBS (pH 7.4) containing 0.1% DMSO as a co-solvent. c Fluorescence intensities of Q-dsAMC (10 mM) after mixing with NADH (0–150 μM), DT-diaphorase (5 μg/mL) in DPBS (pH 7.4) containing 0.1% CHAPS. Error bars represent S.D. (n = 4). d Fluorescence intensities of Q-dsAMC (10 μM) after mixing with d-lactate (0–100 μM), NAD+ (100 μM), DT-diaphorase (5 μg/mL), and d-lactate dehydrogenase (2.5 U/mL) in DPBS (pH 7.4). Error bars represent S.D. (n = 3). e Fluorescence intensities of (left) resazurin (1 μM) or Q-dsAMC (1 μM) after mixing with A549 cells (1 × 105 cells/mL), l-lactate (0 or 100 μM), NAD+ (100 μM), DT-diaphorase (1 U/mL), and lactate dehydrogenase (LDH) (0 or 1 μg/mL) in DPBS (pH 7.4). f Fluorescence intensities of Q-dsAMC (10 μM) after mixing with NAD+ (100 μM), l-lactate or d-lactate (100 μM) with LDH or d-lactate dehydrogenase (DLDH) (5 μg/mL) in PBS (pH 7.4) containing 0.1% CHAPS and incubated for 10 min. The value is mean ± S. D. (n = 4).

Back to article page