Joe98 Posted February 15, 2020 Share Posted February 15, 2020 So I was doing a Bradford assay using spectronic 200 . The standard BSA absorbance I got for 1.00 mg/mL and 1.4 mg/mL was 1.2 and 1.4 which is out of the spectrometer LoL. My lab technician told me that it’s normal. Does anyone know why it’s assumed that we have a linear relationship above an absorbance of 1.00?? Link to comment Share on other sites More sharing options...
BabcockHall Posted February 25, 2020 Share Posted February 25, 2020 My own experience is that the shape of the standard curve depends upon the identity of the protein. However I generally put little trust in absorbance values of greater than 1.0 in the Bradford assay, even using a research-grade instrument, let alone one that is not. The problem is that there is not as much excess Bradford reagent as one moves higher in protein concentration. A standard curve should be made, preferably with the same protein that is being assayed. Link to comment Share on other sites More sharing options...
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