Cell culture methods, transient expression of α1B- and β2-ARs, and membrane preparation

COS-1 and HEK293 cells [American Type Culture Collection (ATCC)] were cultured in Dulbecco’s modified Eagle’s medium containing glutamine and 5 and 10% fetal bovine serum, respectively. CHO-1 cells (ATCC) were cultured in F-12 medium containing 10% fetal bovine serum. Cells were transiently transfected with purified plasmid DNA encoding wild-type or mutant α1B- and β2-ARs, using FuGENE HD in a 1:3 ratio of DNA and FuGENE, following the manufacturer’s protocol. Cell membranes were prepared 48 hours after transfection as described previously for α1B-ARs (41) and prepared the same way for the β2-ARs but in β2 assay buffer [50 mM tris-HCl and 3 mM MgCl2 (pH 7.4)].

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.