Rxn mixtures consist of 5 mM o-AminoBenzaldehyde (oAB) (dissolved in ethanol, & diluted in Glycine-HCl pH 2 buffer) reacted with 20 fold dilution of racemic pyrroline-5-carboxylate (DL-P5C) sample in acidic 1 M HCl diluent. Total volume loaded per well in 96-well plate is 200 ul. Ultimately, absorbances help estimate concentration of DL-P5C in sample, thus I need help determining if 2 hours or longer time duration of days is necessary for sufficient reaction time of oAB-P5C mixtures before taking 443 nm absorbance measurements in plate reader.
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