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Stain glycoprotein

PAS (periodic acid Schiff) stain is used for the histological staining of carbohydrates it is also used to stain glycoproteins (proteins that contain carbohydrates Chap. 2) in electrophoretic gels (Chap. 4). The stain mixture contains periodic acid (HI04), a powerful oxidant, and the dye basic fuchsin ... [Pg.2]

In addition to the methods described above a battery of other staining procedures are available. These include use of alcian blue (22) to stain glycoproteins, ethidium bromide (23) to stain DNA, and methylene blue (14) and pyronine (16) to stain RNA. A relatively new stain has been nicknamed stains-all, because of its ability to stain most macromolecules. This dye is a cationic carbocyanine and stains RNA bluish purple, DNA blue, protein red, acid mucopolysaccharides various shades of blue to purple, and phosphoproteins blue (24). It is presently the most widely used stain for RNA. [Pg.216]

The traditionally minded biochemist stains glycoproteins in SDS gels with PAS. For this, the gel with the periodate-oxidized glycoproteins is successively treated with arsenite and SchifFs reagent (Fairbanks et al. 1971). Gerard (1990) describes a modification of the PAS stain. The PAS stain requires 3 g protein/band and can be characterized by two words inconvenient... [Pg.206]

Figure 52-3. Diagrammatic representation of the major proteins of the membrane of the human red blood cell separated by SDS-PAGE. The bands detected by staining with Coomassie blue are shown in the two left-hand channels, and the glycoproteins detected by staining with periodic acid-Schiff (PAS) reagent are shown in the right-hand channel. (Reproduced, with permission, from Beck WS, Tepper Ri Hemolytic anemias iii membrane disorders, in Hematology, 5th ed. Beck WS [editor]. The MiT Press, 1991.)... Figure 52-3. Diagrammatic representation of the major proteins of the membrane of the human red blood cell separated by SDS-PAGE. The bands detected by staining with Coomassie blue are shown in the two left-hand channels, and the glycoproteins detected by staining with periodic acid-Schiff (PAS) reagent are shown in the right-hand channel. (Reproduced, with permission, from Beck WS, Tepper Ri Hemolytic anemias iii membrane disorders, in Hematology, 5th ed. Beck WS [editor]. The MiT Press, 1991.)...
This process seems to reflect gradual increases in the intensity and density of labelled receptor cell bodies and their axons, followed by regional bulbar staining as synaptogenesis proceeds. However, a more extensive range of lectins needs to be examined before the implications of species differences in the membrane glycoproteins can be satisfactorily interpreted (Salazar and Quinteiro, 1998). [Pg.91]

Attempts to study the entry of ES products into cells using markers of fluid phase endocytosis yielded unexpected results. When larvae browse resistant IEC-6 cells in the presence of extracellular fluorescent dextran, dextran enters the cytoplasm of a significant proportion of the cells in the mono-layer (Butcher et al., 2000). The parameters of dextran entry are most compatible with the conclusion that larvae wound the plasma membranes of IEC-6 cells that is, they create transient breaches in the membrane that allow impermeant markers to enter the cell (McNeil and Ito, 1989). Wounding is considered to be a common occurrence in intestinal epithelia (McNeil and Ito, 1989). Injured cells are able to heal their wounds by recruiting vesicles to seal the breach (Steinhardt et al., 1994). In an experimental system, healing allows the injured cell to retain cytoplasmic dextran. In epithelial cell cultures inoculated with T. spiralis larvae, the relationship between glycoprotein delivery and injury of plasma membranes is not clear, i.e. dextran-laden cells do not always stain with Tyv-specific antibodies and... [Pg.121]

Golgi, in early neuroanatomical studies (1898) staining neurones by silver impregnation, observed a reticular apparatus which was crescent shaped and appeared to be linked through canaliculi. The structure was also seen in secretory cells. Between 1949 and 1954, Baker reported the presence of similar systems in unfixed cells examined by phase contrast. The structures could be stained by osmium tetroxide and probably contained lipid. They also stained for glycoprotein and alkaline phosphatase. Baker s confirmation of the existence of the... [Pg.154]

Glycoprotein Carbohydrate Analysis. Gels from PAGE were fixed with iso-propanol-acetic acid-water (25 10 65 v v v), stained with 0.2% thymol in the fixing solution, and washed two or three times with the same fixing solution. This was followed by staining with concentrated H2S04-ethanol (4 1 v/v) at 35°C for 2-3 h (72). [Pg.418]

This procedure is not solely specific for carbohydrate side chains of proteins. Unglycosylated proteins may also be stained. To identify glycosylated proteins, the sample should be run in at least two identical lanes cut the gel and stain a lane with the common protein silver stain (Protocols 2.4.2.1 to 2.4.2.4) and the other lane by the described method. Compare pattern and intensity to identify glycoproteins. Glycosylated macromolecules are also stainable with Schiff s reagent (Protocol 2.4.4.1), but with less sensitivity. [Pg.60]

PAS staining (periodic acid - Schiff s reagent) colors compounds with vicinal hydroxyl groups, i.e., mainly oligosaccharide side chains in glycoproteins, glycolipids, and nucleic acids. The sensitivity and stability is much lower than the silver staining Protocol 2.4.2.S but more specific. [Pg.62]

When no colored molar mass markers of special glycoprotein markers are used, the ANS staining is recommended, because the geometry of the gel is altered during the manipulations and comparison with Coomassie or otherwise stained gels is difficult. [Pg.63]


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See also in sourсe #XX -- [ Pg.206 , Pg.207 ]




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