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Digitale Medien

Electro-immunoblotting characterization ofPseudo-nitzschia multiseries andP. pungens antigens recognized by antibodies directed against whole cells (1996)

Ross, Neil W. ; Bates, Stephen S.

Springer

Journal of applied phycology 8 (1996), S. 51-58

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ISSN: 1573-5176

Schlagwort(e): Amnesic shellfish poison ; domoic acid ; Pseudo-nitzschia multiseries ; Pseudo-nitzschia pungens ; polyclonal antibody ; Western blot

Quelle: Springer Online Journal Archives 1860-2000

Thema: Biologie

Notizen: Abstract Separate polyclonal antibodies have previously been developed against the domoic-acid-producingPseudonitzschia multiseries (=Pseudo-nitzschia pungens f.multiseries) and the non-toxicP. pungens (=P. pungens f.pungens). These antibodies bind to the surface of the diatoms as shown by immunofluorescence studies. Here we examine the molecular nature of the antigens by Western blotting (electro-immunoblotting) analysis. The major antigens for both polyclonal antibodies migrated as high molecular-weight diffuse bands, mostly remaining in the stacking gel, using an SDS-PAGE system. The antibodies prepared againstP. multiseries strongly labelled the high molecular-weight antigens of allP. multiseries strains tested and showed little reactivity towardsP. pungens extracts on Western blots.P. pungens antibodies strongly labelled high molecular-weightP. pungens antigens and faintly labelled a fewP. multiseries antigens. The selectivity of the antibodies for their respective species correlates with the results of the immunofluorescence experiments, suggesting that the antigens examined in this study are responsible for the selective labelling in immunofluorescence studies. The electrophoretic mobility and the antibody labelling of antigens were not altered by proteolytic digestion of cell pellets. However, disruption of carbohydrates in the pellets by treatment with periodic acid resulted in loss of the antigen. These data suggest that the major antigens of toxicP. multiseries and non-toxicP. pungens are high molecular-weight (°〉100kDa) polysaccharides located on the surface of these diatoms.

Materialart: Digitale Medien

URL: http://dx.doi.org/10.1007/BF02186222

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Digitale Medien

A silver stain protocol for proteins yielding high resolution and transparent background in sodium dodecyl sulfate-polyacrylamide gels (1995)

Swain, Marianne ; Ross, Neil W.

Weinheim : Wiley-Blackwell

Electrophoresis 16 (1995), S. 948-951

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ISSN: 0173-0835

Schlagwort(e): Silver staining ; Protein ; Sodium dodecyl sulfate-polyacryl-amide gel electrophoresis ; Chemistry ; Biochemistry and Biotechnology

Quelle: Wiley InterScience Backfile Collection 1832-2000

Thema: Biologie , Chemie und Pharmazie

Notizen: A simple, sensitive silver staining method for sodium dodecyl sulfate polyacrylamide gel electrophoresis has been developed which yields high resolution of proteins with a transparent background. The method includes the incorporation of a water wash after a short fixation step in ethanol/acetic acid and prior to glutaraldehyde cross-linking, which appears to be necessary for the high resolution staining of protein bands and the low background staining.

Zusätzliches Material: 2 Ill.