Cell Proliferation Kit I (MTT) Protocol & Troubleshooting

Product No. 11465007001

Protocol

Use with yeast or bacteria

Roche has not used the kit to measure of bacterial or yeast proliferation; the kits have not been developed for this purpose. Probably the simplest way to measure bacterial or yeast proliferation is to determine the OD.

Simultaneous use of Cell Proliferation Reagent WST-1, Cell Proliferation Kit I (MTT) and Cell Proliferation Kit II (XTT)

MTT/XTT/WST-1 assays use different tetrazolium salts to measure metabolic activity in active cells. Each kit requires the different indicated wavelength to detect metabolic activity, MTT (550 nm), XTT (492 nm), and WST-1 (420-480 nm). Signal discrimination for each salt is difficult. Sensitivity and linear detection range of established kits run independently would be compromised using combined salts derived from MTT, XTT, and WST-1. For these reasons, simultaneous use of MTT, XTT, and WST-1 is not recommended.

Time frame for reading the plate

Because solubilized formazan is gradually reoxidized, waiting after addition of the solubilization solution can result in a reduced signal intensity.

Troubleshooting

Solubilization of cells

After the 4-hour incubation step with MTT, do not remove MTT and medium prior to solubilization of cells. Add 100 μl of the solubilization solution directly to wells containing culture medium and MTT.

Materials

     
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