Sucrose+gradient+centrifugation

Basic description:

A basic description technique of sucrose density gradient centrifugation is the technique for proteins purification and particularly nucleic acids and it separates particles solution according to their shape, size, density and rotor speed (Bricker 2010).

Purpose of the technique: The purpose of this technique is to separate particles of the same shape but different size. The particles will begin to separate into residues through the gradient according to their shape and density (Bricker 2010).

Origin and History:

The first centrifuge was developed in 1800s by Antonin Prandtl. In 1864, Antonin Prandtl invented the centrifuge to separate cream from milk. His centrifugal method consisted of an upright shaft arms to hold bucket which will increase of the bucket at a certain angle. The machine was running about 400 resolutions per minutes for 15 to 18 minutes (The University of California 2009). In 1920s, T. Svedberg and J.W. Williams developed ultracentrifuge; their technique proved that protein were larger molecules that could weigh in a centrifuge (Bricker 2010). And in 1940s, there was a commercial development of a centrifuge instrument known as the preparative ultracentrifuge; it was available to separate cell in high speed rotation (Alberts et al., 2008, p. 510). Recent Research:

The first recent research using sucrose density gradient centrifugation is “Evidence of copurification of micronuclei in sucrose density gradient-enriched plasma membranes from cell lines” analyzed how centrifugation separated the proteins enriched plasma membrane, and an important of this technique revealed that centrifuging plasma membrane enrichment of micronuclei due to the densities or membrane vesicles (Damarajua et al. 2010). The second recent research “Evaluation of liposome populations using a sucrose density gradient centrifugation approach coupled to a continuous flow system” was also done by using sucrose density gradient centrifugation. In this article, they use sucrose density gradient centrifugation technique to separate liposomes according to the type of liposome, size, and polydispersity. This article revealed that this technique is useful to separate liposomes according to their sedimentation rate of liposome size and density (Sanchez-Lopez et al. 2009). Finally the final research paper, this paper analyzed that when using sucrose gradient centrifugation the major component of microvesicles such a MFG-E8, was clearly detected and the amount in colostrum-derived pellet was higher as well (Hata et al. 2010).

References: Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K., Walter, P. (2008). Molecular Biology of the Cell (5th ed.). //New York: Garland Science//. p. 510; 1288-1290. Bricker, J.V. (2010, Jan 21). //Theory of Ultracentrifugation: Svedberg Equation.// Retrieved from http://bricker.tcnj.edu/tech/BIOL311centrifugation.html Damarajua, S., Zhanga, N., Lic, N., Taoc, L., Damarajud, V.L., Dufoura, J., Santosd, C., Sund, X., Mackeya, J., Wisharta, D.S., Cassa, C.E., Li, L. 2010. Evidence of copurification of micronuclei in sucrose density gradient-enriched plasma membranes from cell lines. //Analytical Biochemistry//, 396, 69–75. Hata, T., Murakami, K., Nakatani, H., Yamamoto, Y., Matsuda, T., Aoki, N. (2010). Isolation of bovine milk-derived microvesicles carrying mRNAs and microRNAs. Biochemical and Biophysical Research Communications, 296, 528-533. The University of California (2009, June 15). //Creamery Industry.// Retrieved from http://books.google.com/books?id=fncSAQAAIAAJ&dq=antonin+prandtl+ centrifuge&source=gbs_navlinks_s