The MTT assay was performed as described in David Morgan's “Polyamine Protocols. Methods in Molecular Biology”33 with minor changes of the protocol as described: 24 h after plating, 50% confluent cells were treated in concentration series including the expected IC50 concentration of the respective substance. After 72 h of incubation the MTT reaction and photometric analysis was implemented. The cell culture medium was aspirated with a vacuum pipette before applying 100 µL MTT working solution (MTT stock solution: 20 mg/ml MTT (Sigma, Cat. No. M5655) in 1× PBS (D8537, Sigma); MTT working solution: 1:5-dilution of the stock solution with RPMI 1640 medium without phenol red). The reaction was stopped after 2 h at 37 °C with 100 µL isopropanol (70 %, Cat. No. 3889017). The photometric analysis at λ = 560 nm was performed in a TECAN Plate Reader after an additional 30 min incubation at room temperature (RT). The viabilities were normalized to DMSO controls. For the identification of the IC50 values all experiments were performed in three replicates and repeated in at least three independent experiments.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.