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The instruction book warns against pyruvate in samples causing background. I also tested the assay on pure oxaloacetate solution and saw a positive reading suggesting that this assay can react non-specifically with oxaloacetate. I believe this assay to not be effective in determining specifically the alphaketoglutarate concentration in samples. I would welcome any advice or suggestions.
Asked on Feb 14 2013
Thank you for contacting us. Did you compare the signal you obtained from the oxaloacetate with the results from a negative control? The control we recommend is testing without addition of the converting enzyme, in order to account for signal contributed by pyruvate and other probe reactants, and subtracting this value from the values obtained from the samples that are incubated with the converting enzyme.
Answered on Feb 14 2013