Deidentified blood was obtained from healthy donors [University of New Mexico (UNM) Hospital Blood and Tissue Bank and Mini Donor Dienst UMC Utrecht], and PBMCs were isolated by Ficoll-Paque gradient separation (GE Healthcare). PBMCs were allowed to rest for 1 hour in RPMI 1640–1% FCS at 37°C in (uncoated) eight-well Lab-Tek chambers. Next, PBMCs were stimulated with human TNFα and IFNγ (500 and 400 U/ml, respectively) for 1 hour in RPMI 1640–1% FCS at 37°C. During the last 10 min of incubation, IV.3 antigen binding fragments [F(ab′)2] (5 μg/ml) and 3G8 F(ab′)2 (1 μg/ml) were added to block the other FcγR (40). Wells were washed with RPMI 1640–1% FCS to remove all unbound cells, leaving the adherent cells (monocytes) in the wells. Next, monocytes were stained for SPT or super-resolution imaging.

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.