I'm setting up a liquid handling robot and need a simple assay to test it. Ideally I would like to mix 3-4 reagents and have a absorbance/fluorescence readout on a plate reader. It's also possible to vary the temperature. Any suggestions?
Kindly check this useful material https://www.ncbi.nlm.nih.gov/books/NBK343429/
1. If the plate reader can read at UV wavelengths, one simple, cheap assay is to monitor the consumption of hydrogen peroxide by catalase at 240 nm.
You just need to mix hydrogen peroxide, a catalase source (it can be a droplet of blood, or a chicken liver homogenate), and phosphate buffer. If using blood, remember to lyse red blood cells, using distilled water, for example.
Final concentrations for good measurements are: 10 mM H2O2 and 50 mM Phosphate buffer (pH 7.0-7.2). Depending on the catalase source, you need to test several concentrations/dilutions.
2. If you do not want to use any biological material, you can do something similar by mixing hydrogen peroxide and Fe(II) and monitoring the absorbance at 240 nm.
Depending on the settings for your demo (is waste disposal an issue?) you could simply use dilutions of food colorants from your local supermarket. If you want a reaction that is producing changes in colors, maybe go for iron(III) (dissolve a nail in dilute hydrochloric acid and expose to air for some time) with rhodanide (isothiocyanate), which will yield a deep red, blood like, color, or use a pH indicator dye (synthetic or e.g. red cabbage extract) with some vinegar, citric acid or soda.
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