A Sendai virus-induced transcriptase-template complex was isolated from the cytoplasm of infected cells by combined sedimentation and isopycnic centrifugation. This transcriptive complex banded at 1.27 g/cm(3) in D(2)O-sucrose gradients. It contained two polypeptides, the viral nucleocapsid structure unit (molecular weight, 60,000) and the largest virion polypeptide (molecular weight, 75,000). ...

• Modern techniques for extracellular vesicle isolation and separation are reviewed. Emphasis is given to modern size-, charged-, affinity-based approaches. A trend toward the combination of multiple EV isolation. Studies subpopulations require combinations techniques. Automated on-line technique allows quick reliable subpopulation studies. Extracellular vesicles (EVs) heterogenous membrane-bou...

Intercellular communication can be carried out by circulating systemic and/or locally released extracellular vesicles (EVs), produced nearly every cell type and tissue, are involved in physiological pathological processes. In recent years, EVs have been identified reproductive tissues, such as oviduct uterus, shown to related several events important for success. The understanding of their func...

Extracellular vesicles (EVs) are released by mammalian cells and are thought to be important mediators of intercellular communication. There are many methods for isolating EVs from cell culture media, but the most popular methods involve purification based on ultracentrifugation . Here, we provide a detailed protocol for isolating EVs by differential ultracentrifugation and analyzing EV protein...

Using a commercially available test for LDL cholesterol (Boehringer Mannheim), a method was developed for determination of LDL apolipoprotein B without using ultracentrifugation. The infranatant obtained by precipitation of serum with polyvinylsulphate was redissolved in a saline citrate solution and incubated with phospholipase A2, phospholipase C, phospholipase D or triglyceride lipase, respe...

Retroviral and lentiviral vectors are mostly pseudotyped and often purified and concentrated via ultracentrifugation. In this study, we quantified and compared the stabilities of retroviral [murine leukemia virus (MLV)-based] and lentiviral [human immunodeficiency virus (HIV)-1-based] vectors pseudotyped with relatively mechanically stable envelope proteins, vesicular stomatitis virus glycoprot...

We evaluated the density-gradient ultracentrifugation method in a swinging-bucket rotor (Anal Biochem 111, 149-157, 1981) in a slightly modified version for isolation and determination of high-density lipoprotein (HDL) subfractions. We prestained the serum with Coomassie Brilliant Blue R, which did not change the hydrated densities of the lipoproteins, and after only 2.2 X 10(8) gav . min obtai...