Treated Hela cells on coverslips were washed once with prewarmed PBS and fixed in 4% (w/v) paraformaldehyde for 15 min. After three washes in PBS, cells were permeabilized with 0.2% (v/v) Triton X-100 for 5 min. After three washes in PBS, cells were blocked in PBS containing 5% (w/v) bovine serum albumin (BSA) for 30 min and incubated with indicated antibodies in PBS containing 3% (w/v) BSA for 2 hours at 37°C. After three washes, cells were incubated with Alexa Fluor 488–conjugated secondary antibodies for 1 hour at 37°C and then with 4′,6-diamidino-2-phenylindole (Roche) for 15 min. The coverslips were washed extensively and mounted onto slides. Imaging of the cells was carried out using an Olympus BX51 microscope confocal system under a 40× objective.

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

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.