RNA polymerase II from larvae of the brine shrimp, Artemia salina, was highly purified by two cycles of DEAE-cellulose chromatography followed by centrifugation through discontinuous sucrose gradients. Gradient fractions were subjected to elctrophoresis is polyacrylamide gels containing sodium dodecyl sulfate. The subunit structure of RNA polymerase II was determined by quantitative comparison ...

Despite numerous studies, isolating pure preparations of extracellular vesicles (EVs) has proven challenging. Here, we compared ion-exchange chromatography (IEC) to the widely used sucrose density gradient (SDG) centrifugation method for the purification of EVs. EVs in bulk were isolated from pooled normal human amniotic fluid (AF) by differential centrifugation followed by IEC or sucrose densi...

Preparation of the nanodiamonds (NDs) The raw, detonated NDs (NanoCarbon Institute, Japan) were acquired as colloidal particles and then dispersed in deionized water. The raw ND colloid was carboxylated in strong oxidizing conditions as previously reported. Briefly, the colloid was treated in acid mixture of 68% HNO3 : 98% H2SO4 ( 1:3 in volumn ) in ultrasonic bath for 24 h, followed by stirrin...

This study reports on the distribution of bicarbonate-stimulated ATPase in rat intestinal epithelial cells. Brush-border membranes and basolateral membranes were separated from each other and from mitochondrial and other intracellular membranes by differential and density gradient centrifugation. Bicarbonate-sensitive ATPase activity followed the mitochondrial marker succinic dehydrogenase clos...

Intracellular localization of lunularic acid and prelunularic acid in suspension cultured cells of Marchantia polymorpha L. was studied. The sum of both compounds was determined as lunularic acid group (LNAs) because of the instability of prelunularic acid to convert into lunularic acid.Mechanical disruption of the cells followed by differential centrifugation showed that LNAs was associated wi...

In this paper, we describe a cultivation procedure in large-capacity fermentors (200 liters) and the purification of the Salmonella crude extracts by means of zonal centrifugation in a sucrose gradient. Briefly, the endotoxin has been extracted from salmonellae with hypertonic solutions. The crude endotoxin was shown to contain several antigens, mainly r(1), r(2), r(3), or heterologous antigen....

Background: Techniques of sperm preparation require centrifugation to separate spermatozoa from the seminal plasma. The centrifugation can induce damage to the spermatozoa and increase reactive oxygen species(ROS) productionspecially in leukospermic speciemens that are associated with impaired function and fertilizing capacity. However in ICSI, fertilization can occure with damage DNA sperm but...