A highly sensitive periodic acid-silver stain for 1,2-diol groups of glycoproteins and polysaccharides in polyacrylamide gels

Dubray, Gérard; Bezard, G.

doi:10.1016/0003-2697(82)90593-0

Cited by 339 publications

(168 citation statements)

References 11 publications

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“…Wells were loaded with 7 µg of protein. The resulting protein patterns were stained with a combined Coomassie Blue-Silver staining procedure (De Moreno et al 1985) for proteins and combined Periodic Acid-Alcian Blue-Glutaraldehyde-Silver (PAABGS) stain procedure for glycoproteins (Dubray & Bezard 1982, Moller et al 1993, Moller & Poulsen 1995. For glycoproteins, two types of markers were run in the same gel: the broad molecular weight range was used to estimate molecular weight and control the reversal of silver staining (unstaining) of proteins and a mixture of proteoglycans and glycoproteins (6 µg/well).…”

Section: Methodsmentioning

confidence: 99%

Morphological comparison of axenic amastigogenesis of trypomastigotes and metacyclic forms of Trypanosoma cruzi

Navarro

Lima

Askue

et al. 2003

Mem. Inst. Oswaldo Cruz

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Section: Methodsmentioning

confidence: 99%

Morphological comparison of axenic amastigogenesis of trypomastigotes and metacyclic forms of Trypanosoma cruzi

Navarro

Lima

Askue

et al. 2003

Mem. Inst. Oswaldo Cruz

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“…Antibody binding was detected using goat antirabbit HRP-conjugated antibodies (BioRad) and an ECL kit (Amersham, Buckingham, UK). Periodate-Schiff reagent (PAS) staining was performed as described previously [9]. Spectroscopy was performed using a Novaspec II spectrophotometer (Pharmacia, Uppsula, Sweden).…”

Section: Other Methodsmentioning

confidence: 99%

Characterization of Inverted Membrane Vesicles from the Halophilic Archaeon Haloferax volcanii

Ring

Eichler

2001

Journal of Membrane Biology

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Abstract. Membrane-related processes in archaea, the third and most-recently described domain of life, are in general only poorly understood. One obstacle to a functional understanding of archaeal membrane-associated activities corresponds to a lack of archaeal model membrane systems. In the following, characterization of inverted archaeal membrane vesicles, prepared from the halophilic archaeon Haloferax volcanii, is presented. The inverted topology of the vesicles was revealed by defining the orientation of membrane-bound enzymes that in intact cells normally face the cytoplasm or of other protein markers, known to face the exterior medium in intact cells. Electron microscopy, protease protection assays and lectin-binding experiments confirmed the sealed nature of the vesicles. Upon alkalinization of the external medium, the vesicles were able to generate ATP, reflecting the functional nature of the membrane preparation. The availability of preparative scale amounts of inverted archaeal membrane vesicles provides a platform for the study of various membranerelated phenomena in archaea.

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“…The blots were washed and visualized by using horseradish peroxidase-conjugated goat anti-mouse immunoglobulin G antibody (Sigma Chemical Co., St. Louis, Mo.) as described above or a silver stain protocol identical to that described by Dubray and Bezard (15).…”

Section: Methodsmentioning

confidence: 99%

Heterogeneity of lipopolysaccharides from Pseudomonas aeruginosa: analysis of lipopolysaccharide chain length

et al. 1988

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Lipopolysaccharide (LPS) from smooth strains of Pseudomonas aeruginosa 503, PAZ1, PA01715, PA01716, and Z61 was fractionated by gel filtration chromatography. LPS samples from the first four strains, all PAO1 derivatives, separated into three major size populations, whereas LPS from strain Z61, a Pac K799/WT mutant strain, separated into two size populations. When column fractions were applied to sodium dodecyl sulfate-polyacrylamide gels in their order of elution, molecules of decreasing size were resolved, and the ladder of molecules with different-length 0 antigens formed a diagonal across the gel. The LPS from the PAO1 derivatives contained two distinct sets of bands, distinguished on the gels as two sets of diagonals. The set of bands with the faster mobility, the B bands, was found in column fractions comprising the three major amino sugar-containing peaks. In the sample from strain 503, a fourth minor peak which contained B bands was resolved. The slower-moving set of bands, the A bands, were recovered in a minor peak. LPS from strain Z61 contained only one set of bands, with the higher-molecular-weight molecules eluting from the column in a volume similar to that of the B bands of the PAO1 strains. Analysis of the fractions of LPS from all strains indicated that less than 8% of the LPS molecules had a long, attached 0 antigen. Analysis of the peak that contained mainly A bands indicated a lack of reactive amino sugar and phosphate, although heptose and 2-keto-3-deoxyoctulosonic acid were detected. Reaction of isolated fractions with monoclonal antibody specific for the PA01 0-antigen side chain indicated that only the B bands from the PAO1 strains were antigenically reactive. The bands from strain Z61 showed no reactivity. The data suggest that the A and B bands from the PAO1 strains are antigenically distinct. We propose that PAO1 strains synthesize two types of molecules that are antigenically different.Lipopolysaccharide (LPS), a major component of the outer membranes of gram-negative bacteria, is important in the structure (33, 37) and function (33,36) of this membrane. Structural microheterogeneity in several regions of LPS molecules from members of the family Enterobacteriaceae (2,14,22,37,38,48,51) and Pseudomonas aeruginosa strains (33, 57) has been demonstrated. Of the several methods used to separate the subclasses of LPS from individual strains, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (20,24,45,48) and gel filtration (9, 26, 30, 32, 34, 48) are the best. Either of these two methods by themselves, however, may be insufficient to completely characterize the high-and low-molecular-weight fractions of LPS. Peterson and McGroarty (48) demonstrated that the SDS-PAGE of the column fractions of samples from Salmonella typhimurium, Salmonella minnesota, and Escherichia coli was instrumental in characterizing the various-sized fractions. Analysis of the isolated fractions allowed for the estimation of the average number of 0-antigen repeat units per LPS from each of the si...

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A highly sensitive periodic acid-silver stain for 1,2-diol groups of glycoproteins and polysaccharides in polyacrylamide gels

Cited by 339 publications

References 11 publications

Morphological comparison of axenic amastigogenesis of trypomastigotes and metacyclic forms of Trypanosoma cruzi

Morphological comparison of axenic amastigogenesis of trypomastigotes and metacyclic forms of Trypanosoma cruzi

Characterization of Inverted Membrane Vesicles from the Halophilic Archaeon Haloferax volcanii

Heterogeneity of lipopolysaccharides from Pseudomonas aeruginosa: analysis of lipopolysaccharide chain length

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