Sucrose density gradient centrifugation

The purification is done by sucrose density-gradient centrifugation (DeSa and Hastings, 1968 Hastings and Dunlap, 1986). Six sucrose... 

The PemB cellular localisation was determined both in E. chrysanthenu and in an E. coli recombinant strain by Western blot of the cell fractions with a PemB-antiserum. No PemB was detected in the culture supernatant and only trace amounts were found in the soluble cell fractions - periplasm and cytoplasm (Figure 2). PemB was found mostly in the total membrane fraction from which it could be completely extracted by Triton X-100/Mg2+ and partially extracted by Sarkosyl (Figure 2). This behaviour is typical of inner membrane proteins, but since some exceptions have been noticed it does not positively indicate the PemB localisation (15). We performed cell membrane fractionation in sucrose density gradient centrifugation both by sedimentation and flotation, using several markers of inner and outer membrane vesicles. PemB was found in the outer membrane vesicles (data not shown). 

Fig. 2. Preparation of linear sucrose density gradient, centrifugation, and fractionation. Fractionation is performed using an ISCO model 640 density gradient fractionator in this experiment.

Fig. 4. Localization of WAP27 and WAP20 in the crude microsome fractions and the relation with marker-enzyme activities in three organelles (ER, tonoplast, and Golgi). SDS-PAGE of fractionated proteins by isopycnic linear sucrose density gradient centrifugation of microsome fraction of mulberry cortical parenchyma cells was performed using 6-pL samples in each fraction. Immunoblot analysis was performed with anti-WAP27 and anti-WAP20 antibodies. (From ref. [1], with permission from the American Society of Plant Physiologists.)...

The molar ratio of the components of metaphase chromosome was determined by Uchiyama and his colleagues (Uchiyama et al., 2005). Metaphase chromosomes has been isolated from synchronized human cell line and then purified by sucrose density gradient centrifugation and Percoll density gradient centrifugation. The major components described in this chapter are summarized here. Molar ratio is provided per 100 histone H4 molecules. 

RNA Separation by Non-Denaturating Sucrose Density Gradient Centrifugation... 

For calculation it was assumed that these five phospholipids together accounted for all of the phospholipids of the membrane. In the actual analyses, other phospholipids (primarily lysophosphatidyl choline and lysophosphatidyl ethanolamine) were found to account for less than 5% of the total. The heavy and light microlipid droplet fractions were obtained by sucrose density gradient centrifugation. Heavy fractions were collected between 0.5 and 1.0 M sucrose and light fractions banded at the 0.5 M sucrose-buffer interface. 

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