Absorbance reading does not match spectra
Posted 06 September 2010 - 01:01 AM
I am using a new substrate so after my assay I ran a spectra (200-800nm) to determine the optimal wavelengh. The spectra looked very good with the peak at 630nm close on 2 absorbance units. I then read the wells at a single wavelengh of 630nm but the readings were in the region of 0.4 Au. Does anyone know why this occurs? And what can I do about it?
Just to add, I read at various wavelengths after that (i.e. 600, 610, 620, 640, 650, 660, 670, 680, 690, 700) as I received the same results - all reading in the range of 0.4 Au.
Any advice on this will be gratefully accepted.
Posted 06 September 2010 - 11:54 AM
The readings from your sample looks as the didn't react, a other possibility is that the reactionproduct breaks down. I,m just guessing by lack of information.
Pluralitas non est ponenda sine necessitate
-- "You must assume no plural without necessity".
Posted 07 September 2010 - 06:19 AM