Purification of the human intestinal brush border membrane

Intestinal Farnesoid X Receptor Controls Transintestinal Cholesterol Excretion in Mice

“…Intestinal brush border membranes were isolated as described by Schmitz et al 29 Equal amounts of brush border membranes (10 µg protein) for detection of ABCG5 were electrophoresed and blotted on Hybond ECL membranes (Amersham, Little Chalfont, UK). Membranes were incubated with anti-ABCG5 primary antibody.…”

Angiotensin I converting enzyme (kininase II) of the brush border of human and swine intestine

“…These are characteristic features of purified brush border vesicles (15) and brush border in situ (9,23). In addition, no significant contamination by other subcellular organelles was apparent.…”

Biosynthesis of microvillar proteins

“…This has undoubtedly to do with the fact that these enzymes, contrary to viral proteins, each represent only a tiny portion (0.1-1%) of the total mass of protein ofthe cell and that their biosynthesis therefore inevitably must be studied against a high background. By using immunofluorescence labelling, Feracci et al (1982) (Schmitz et al, 1973) and kidney (Booth & Kenny, 1974) tissue and has been used in preparative scale in the purification of several microvillar enzymes (Kenny & Maroux, 1982). Using a modification of the procedure of Kessler et al (1978), it was possible to obtain a Ca2 +-precipitated membrane fraction that contains only 2-4% of the total activity of the microvillar enzymes, indicating a minimal contamination with microvillar membranes (Danielsen et al, 1981a).…”

“…In addition, specific antibodies, polyclonal (for references see as well as monoclonal (Hauri et al, 1980;Gee et al, 1983) have been raised against these enzymes. Furthermore, effective methods for isolating microvillar membranes from both intestine and kidney have been described (Schmitz et al, 1973;Booth & Kenny, 1974), as have methods for organ culture of tissue explants for long enough periods to study biosynthetic events (Moscona et al, 1965). Lately, preparations of translatable mRNA from both small intestine (Wacker et al, 1981;Danielsen et al, 1982b) and kidney (Nash & Tate, 1984) have become available, and thus the scientific tools and model systems necessary for a thorough study of the biosynthesis of microvillar proteins are at hand.…”