Hello,
I'm trying to measure trypsin activity using BAEE substrate according to the method from sigma.
https://www.sigmaaldrich.com/technical-documents/protocols/biology/enzymatic-assay-of-trypsin.html
However, the absorbance of BAEE is around 0.6, and the absorbance does not change much after adding trypsin, just around 0.85 in 10min. Hence the activity of trypsin is pretty low. I have no idea about it.
Dear Patrick,
I did not blank BAEE solution, just blank water, what I care about is △A，should I?
I tried again, prepared BAEE solutions with different concentrations, I also diluted my trypsin with different concentrations, and found 0.1mg/ml trypsin was most suitable in this case, presenting a linear change in first 3 min.
Dear Patrick, I calculate " △A/min (with trypsin) --- △A/min (without trypsin)". so I still do not understand why I should blank BAEE solution.
Because many literatures said, the absorbance of BA should dramatically higher than that of BAEE, but in my case, the absorbance of BAEE is around 0.5, and after hydrolysis with trypsin, just up to around 0.7 in 10 min. I'm not sure, is there any problem?
Dear Patrick, thank you, and how about your data when you hydrolyzed BAEE with trypsin?
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