Hi all,
I wonder if there are any tips reg. increasing the assay window?
We perform 4-fold dilutions for 10 points.
Currently in our case, if the initial concentration is relatively high, the upper points will go into plateau, and with low concentrations the lower points form a plateau accordingly. The linear range of increasing concentrations is too small for the assay to work properly (assay window = ~2). Are there any ways to increase this value? We have tried optimizing initial concentration, cell density, or X-fold dilutions, still cannot achieve the assay window increase.
Hi,
Have you tried to use D-optimal design to generate the optimal set of experimental runs? D-optimal design is one forms of design based on a computer-aided exchange procedure to create the optimal set of experiments. Given the total number of treatment runs for an experiment and type of model the experimenter wishes to fit, the computer algorithm generates the optimal set of design runs from all possible combinations of various factor levels that one wishes to use in the experiment. There are computer programs, such as Matlab and SIMCA (MODDE), for generating D-optimal designs for a given experiment.
Please check this link to learn more about D-optimal design
http://www8.cs.umu.se/education/examina/Rapporter/FabianTtriefenbach.pdf
Good luck with your interesting experiment!
Mulualem Tigabu, thanks for your reply! This will be useful designing a development strategy!
Hi Andrej,
depending on the type of your assay, and what you are measuring, you might be successful by trying a different culture medium, a FCS/BCS from a different origin (Australia over South America), prolonged exposure time or choosing a different cell clone less susceptive to compounds triggering the cell response you measure. We have optimized all of these factors successfully with regard to overall induction.
Wishing you best of success
Johannes
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