I am trying to assess the inhibition effect of the thioredoxine reductase by UVvis. The problem is that I have to use a fixed wavelength method within the UV-vis spectrophotometer and as I never did it before I am not sure how to setup the method.
I am using an Evolution 600 spectrophotometer (visionpro software) and within the window of the fixed method I can set 10 different wavelengths. I only have to set up one at 412 nm, as I have to measure the absorbance at that wavelength over time. What I don´t know is that if I have you click on the reference wavelength bottom, and if so, which reference wavelength do I have to place on the bracket. Additionally, is it necessary to do a base line experiment before each measurement.
Apparently visionpro software is not prepared for kinetics measurements (Abs. vs. time).
But, in the attached guide, I see that you can access a kinetics application in local control mode. Read at pages 23 (local control), 25 (main menu), 45 (kinetics application).
The "fixed method" is not a good choice. In any case you may try to activate only one wavelength, no need for reference wavelength correction, use "cycles" at least 200, estimate the experiment duration (e.g., 300 s), choose the "integration time" = experiment duration / "cycles". Autosave = on.
But I recommend to use the local control mode.
Thank you a lot for your help!!! It was very enlightening!!!
kind regards
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