Isolation of Brush-Border Membranes Brush-Border Membranes (BBM) were isolated from frozen jejunal mucosal scrapings using a combination of cation precipitation and differential centrifugation
نویسندگان
چکیده
منابع مشابه
Simple method of preparation of rat colonocyte apical membranes using polyethylene glycol precipitation.
BACKGROUND Although a number of methods are available for the preparation of brush border membranes (BBM) from the small intestine, very few studies have been performed on the isolation of colonic apical membranes (CAM). Cation precipitation is one of the methods used for the isolation of these membranes, which can result in altered lipid composition as a result of activation of phospholipase. ...
متن کاملSimultaneous isolation and characterization of brush border and basolateral membrane vesicles from bovine small intestine.
Purified brush border and basolateral membranes were isolated from homogenized intestinal enterocytes of Holstein steers by divalent cation precipitation followed by differential and sucrose density gradient centrifugation. Alkaline phosphatase and Na/K adenosine triphosphatase served as marker enzymes for the brush border and basolateral membranes, respectively. The brush border and basolatera...
متن کاملAnalytical subcellular fractionation studies on enterocytes from the jejunum and ileum of the rat and some properties of brush-border alkaline phosphatase.
1. Enterocytes, isolated from the proximal jejinum and distal ileum of the rat, were homogenized and their organelles separated by isopycnic centrifugation on continuous sucrose density gradients. The distributions of marker enzymes for the principal organelles, RNA and protein were determined in the sucrose gradients and related to the activities per entercocyte. 2. In the jejunum the modal eq...
متن کاملA simple method of rat renal brush border membrane preparation using polyethylene glycol precipitation.
A simple method for preparation of brush border membranes (BBM) from rat kidney using polyethylene glycol (PEG) precipitation has been described. This method avoids the use of cations for the preparation, which might alter membrane lipid composition. These preparations were assessed for enrichment of marker enzymes, contamination by subcellular structures, lipid composition and transport functi...
متن کاملPreparation of Microvilli from Rabbit Kidney
A simple method for the isolation of microvilli from kidney brush border is described. The method depends on the preferential aggregation of other subcellular structures by bivalent metal ions. MgCl2 is added to a homogenate of cortical tissue prepared from frozen rabbit kidneys. Aggregated material is removed by a low-speed centrifugation and the supematant centrifuged at 15000g to yield a pel...
متن کامل