Extracellular vesicles were isolated according to standard protocols from our group [26 (link), 27 (link)]. Briefly, plasma aliquots were thawed and each 500 µl were diluted into 1 mL of 1X PBS and centrifuged at 3,000 × g for 20 min at 4 °C and later at 10,000 × g for 30 min at 4 °C to remove cell debris. Supernatants were recovered, diluted with 1X PBS, and centrifuged at 100,000 × g for 70 min at 4 °C into 6.5 mL, Open-Top Thickwall Polycarbonate Tubes (Beckman Coulter) in an Optima MAX Ultracentrifuge (Beckman Coulter). Finally, EVs pellets were resuspended in 1X PBS or lysed with 1X RIPA lysis buffer (Cell Signaling) and sonicated for 2 min for further analysis.
de Miguel-Perez D., Russo A., Arrieta O., Ak M., Barron F., Gunasekaran M., Mamindla P., Lara-Mejia L., Peterson C.B., Er M.E., Peddagangireddy V., Buemi F., Cooper B., Manca P., Lapidus R.G., Hsia R.C., Cardona A.F., Naing A., Kaushal S., Hirsch F.R., Mack P.C., Serrano M.J., Adamo V., Colen R.R, & Rolfo C. (2022). Extracellular vesicle PD-L1 dynamics predict durable response to immune-checkpoint inhibitors and survival in patients with non-small cell lung cancer. Journal of Experimental & Clinical Cancer Research : CR, 41, 186.
Other organizations :
Icahn School of Medicine at Mount Sinai, University of Maryland, Baltimore, Instituto Nacional de Cancerología, University of Pittsburgh, University of Pittsburgh Medical Center, UPMC Hillman Cancer Center, The University of Texas MD Anderson Cancer Center, University of Messina, Fondazione IRCCS Istituto Nazionale dei Tumori, Foundation for Clinical and Applied Cancer Research, Universidad El Bosque, Pfizer-University of Granada-Junta de Andalucía Centre for Genomics and Oncological Research
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to
get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
