Small-molecule screening was performed using the ICCB-Longwood Screening Facility (https://iccb.med.harvard.edu). GFP-expressing cells were seeded into black-sided 384-well plates at 600 cells per well using a Multidrop Combi Reagent Dispenser (Thermo Fisher Scientific, no. 5840300) in a final volume of 30 μl per well. After 24 hours, a Seiko Compound Transfer Robot was used to pin transfer 100 nl of each library plate well into the cell-containing plates. Forty-eight hours later, the GFP signal was measured as a proxy for cell number using an Acumen Laser Scanning Cytometer. The Ludwig Anti-Cancer Library of compounds was used and contains ~400 compounds in 10-point concentration curves ranging from 5 nM to 33.3 μM. The average Z′ value for this screening setup was 0.75 when using actinomycin D as the positive control.

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.