HEK293T cells were cultured in 96-well plates in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C in 5% CO2 and transfected with 200 ng of DNA by Lipofectamine LTX (Life Technologies) according to the manufacturer’s protocol. Antigen expression was detected as we described previously (43, 67). Briefly, at 48 hours after transfection, transfected cells were fixed with 4% formalin (Sigma), permeabilized with methanol at −20°C, and then blocked in 2.5% bovine serum albumin (Sigma) in phosphate-buffered saline (PBS) before the addition of anti-NS1 antibody (BioFront Technologies) at 37°C for 2 hours. Finally, cells were stained with DAPI (4′,6′-diamidino-2-phenylindole; Life Technologies) and visualized by fluorescence microscopy (Zeiss LSM-700), and the data were digitized using the Zen software (Zeiss).

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.