What is needed for RapidFire® quantitation of substrate and product peaks?
As part of the RapidFire® Assay development Service we ask that you provide us with a small sample of pure substrate and (if available) product. Using these, we are able to determine the best analytical conditions to use. Briefly, we need to know the MRM transitions to use to detect and quantitate each of the molecules. That is, the mass of each molecule and the mass of a fragment of that molecule that would be generated in the triple-quadrupole mass spectrometer. Generally our clients are able to tell us what the substrate and product molecules are so we determine MRM transitions either from this information, or empirically, using the samples provided to us. Once we know which MRM transitions to track, we monitor these same ones for each well as a measure of the amounts of substrate and product (and often internal standard) present. We never measure the test compounds in the wells.
