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Gradient-forming vessels Al and A2 are 27 mm ID and 200 mm high. The bottom of the gradient former should be higher than the top of the separating chamber (C). The "Rotaflo" tap (R) controls the rate of flow from the gradient. The all glass components are connected by sterilizable silicone tubing. The cell chamber (B) is 18 mm ID and 100 mm ...
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Thus, DRMs can be isolated by flotation as low-density membranes in density gradients, and this constitutes the basis for the original and now widely used operational definition of lipid raft as a membrane domain that floats like a raft on top of a density gradient as a separable, cholesterol-enriched fraction [38,39,40].
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Figure 1. Purification of plastoglobules by flotation centrifugation. Total membranes from isolated chloroplasts were separated by flotation on a continuous sucrose gradient. Plastoglobules are visible as a yellowishgreen layer at the top of the gradient. THY, thylakoid membranes; PG, plastoglobules. Recipes . 1. HB buffer
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By modifying sucrose density gradient flotation centrifugation, which is used to isolate lipid microdomains, we were able to separate caveolae and noncaveolar lipid microdomains into two distinct fractions. The caveolar membranes are membrane vesicles of 100-nm diameter, enriched with caveolin-1 and flotillin-1. ...
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Flotation of nuclei appeared to depend on the low salt concentration used in the bottom of the gradient. We hypothesize that under these conditions, chromatin may at least partially decondense ( 30 ), thus increasing the buoyancy of the structure.
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Flotation methods use a liquid suspending medium that is denser than the oocysts being concentrated. This can be used for the purpose of both concentration and purification. ... The simple or multistep gradient purification strategies to isolate oocysts from contaminants are: SSF, SSSF, ZSF, percoll, glass bead column (GBC), dialysis, cesium ...
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To analyze the role of lipid-rich membranes in HCV RNA replication, we utilized a membrane flotation assay based on an 10-20-30% iodixanol density gradient developed by …
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Flotation with a step sucrose gradient. i. Adjust sample composition to 40% (wt/wt) sucrose by adding 2 ml 56% sucrose. ii. Transfer to a SW40 centrifuge tube and carefully overlay it with 8.5 ml ...
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6. Put the gradient mixer on top of a stir plate so that the gradient maker is level. Put a small stir bar in mixing chamber. 7. With all valves closed, fill each chamber with 1/2 the total amount of desired gradient [i.e. 1.1 ml for a 2.2 ml gradient (TLS-55)]. Use 40% sucrose in mixing chamber and 10% sucrose in reservoir chamber. 8.
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The invention discloses a gradient flotation method for low-activity pyrites, which comprises the following steps of: firstly, only using trace amounts of xanthate, such as butyl xanthate, with usage amount of 15-30g/t, to collect the pyrites with better flotability and high flotation rate, and quickly carrying out flotation to obtain concentrates as high-sulfur concentrates; next, using small ...
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Download scientific diagram | Isolation of extracellular vesicles from plasma. (A) Gradient flotation principle. Plasma samples are density-adjusted from publication: Plasma-derived extracellular ...
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1. Cells were dispersed from human foreskin by proteolytic digestion and enriched or depleted in mast cell content by density gradient flotation on discontinuous gradients of Percoll. 2. Cells were harvested at six interfaces on the density gradient. Mast cell purity ranged from 0.6 …
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Here, we describe a detailed protocol for density gradient fractionation of various mammalian subcellular vesicles, including endoplasmic reticulum (ER), Golgi apparatus, endosomes, and …
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By modifying sucrose density gradient flotation centrifugation, which is used to isolate lipid microdomains, we were able to separate caveolae and noncaveolar lipid microdomains into two distinct ...
WhatsApp: +86 18221755073
membranes in the life cycle of HCV. To analyze the role of lipid-rich membranes in HCV RNA replication, we utilized a membrane flotation assay based on an 10-20-30% iodixanol density …
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3.3. Density gradient fractionation of plasma lipoproteins. Analytical UTC with density gradient flotation using Schlieren optics was used over 70 years ago to characterize lipoproteins . With sequential flotation, lipoproteins could be separated into five major groups such as HDL, LDL, IDL, VLDL and chylomicrons.
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1. With a stationary rotor the AFH loads the gradient. 2. Slow acceleration reorients the gradient. 3. At 35 000 rpm flow sample into the rotor; vector is captured in the density gradient, waste flows out. 4. Isopycnic sedimentation occurs in zones where the gradient density equals a particle's buoyant density. 5.
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Sucrose gradient centrifugation (SGC) based on the different flotation densities could improve the purity. Hogan et al confirmed that SGC can reduce the interference from the amount of Tamm-Horsfall protein in urine [ 32 ], which is a typical contaminant in …
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The methods used for the density gradient flotation of serum lipoproteins follow, in general, those described in our earlier publications [4,5]. We have, however, made a few changes, some due to greater experience and others to harmonise with the methods of Lindgren et al. [6]. Linear density gradients ranging from a density of 1.003 to 1.2 and ...
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Faecal samples from asymptomatic dairy cows and calves from a farm on the Island Falster, Denmark, were examined by a sucrose gradient flotation technique. Giardia cysts were found in 7.6% of the 92 samples, and estimated cyst excretion rates ranged from 50-200 cysts per gram faeces. Given that Giar …
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The U.S. Bureau of Mines has investigated new methods for increasing the selectivity in flotation by creating and regulating reagent concentration gradients within flotation columns at steady-state operation. Gradients were generated and traced in a water-only system with the use of fluorescent dyes, and the effects of various operating parameters on these …
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tive separation by arresting the flotation rate of the lym-phocytes more than that of the monocytes. For a detailed protocol and references see C9 on the OptiPrep™ Application CD or online at: The efficacy of the OptiPrep™ flotation method relies on the slightly lower density of monocytes in combina-
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The aim of this study was to examine a modified flotation density gradient centrifugation (DGC) technique in which semen was diluted with a colloid solution (Opti-prep TM) to increase its density prior to layering between colloid layers of lower and higher density.
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Here we used simple sucrose gradient flotation centrifugation method to purify the plastoglobules from total chloroplast membranes (Vidi et al., 2007, Shanmugabalaji et al., 2013).
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To analyze the role of lipid-rich membranes in HCV RNA replication, we utilized a membrane flotation assay based on an 10–20–30% iodixanol density gradient developed by Yeaman et …
WhatsApp: +86 18221755073
Flotation often provides better recovery and/or resolu-tion of the least dense components of a crude fraction than does sedimentation. Overloading the gradient (instability of the sample/gradient interface) cannot oc-cur in a flotation format. The particular format de-scribed in this protocol also allows the sample to be
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By modifying sucrose density gradient flotation centrifugation, which is used to isolate lipid microdomains, we were able to separate caveolae and noncaveolar lipid microdomains into …
WhatsApp: +86 18221755073
Immunomagnetic separation after sucrose gradient flotation gave superior results compared to sucrose gradient flotation alone. The method enables detailed analysis of a wide range of genes of interest for genotyping, virulence and drug resistance.
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