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  • 1
    ISSN: 1573-4919
    Keywords: Hemagglutinin-filamentous ; gel electrofocusing ; gel electrophoresis-quantitative ; pertussis ; SDS-polyacrylamide gel electrophoresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Summary A highly purified preparation of filamentous hemagglutinin (FHA) from Bordetella pertussis was analyzed for its protein composition by gel electrophoretic methods. In this preparation of FHA the following native species could be detected by polyacrylamide gel electrophoresis (PAGE) at pH 3.2: S, and S2 (inactive subunits or fragments); two monomers, a major form designated Ia (144K), and a minor form lb, differing only in net charge; and three oligomeric forms, designated II (213K), III (595K) and IV (1064K). Hemagglutinating activity was associated predominantly with component Ia. PAGE of FHA after derivatization with sodium dodecyl sulfate (SDS) showed there to be three major species, designated A, C and D. According to estimated molecular weight values, A, C and D are likely to correspond to S2, Ia and II respectively. Isolated components II, III and IV yield all three SDS-species upon derivatization with SDS. Both moving boundary electrophoresis and gel electrofocusing showed hemagglutinating FHA to be a basic protein. Its apparent pI is 8.1.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A device for rapid rotation of perforated gel holding tubes has been constructed by which the diffusion of sodium dodecyl sulfate (SDS) from cylindrical polyacrylamide gels can be substantially accelerated. This device allows one to stain protein bands obtained in SDS-polyacrylamide gel electrophoresis (9 % gel concentration) within one and a half hours, using the procedure of fixation in trichloroacetic acid followed by Coomassie Brilliant Blue G-250 staining, which requires no destaining.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 1 (1980), S. 23-27 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: In isoelectric focusing on polyacrylamide gel, pH gradients exhibited a systematic shift along the pH-axis in proportion to the pH of the anolyte. Thus, more acidic anolytes produced more acidic pH gradients, and more basic ones gave rise to more basic pH gradients. In all cases the pH's of the anolyte and catholyte were kept outside the pI-range of the carrier ampholytes used. Increasing the pH of the anolyte stabilized pH gradients, while the pH of the catholyte, within the range tested, had no systematic effect on pH gradient stabilization.From a practical viewpoint, modification of pH gradients by the anolyte provides increased flexibility in pH gradient design for electrofocusing. From a theoretical viewpoint, the selective anolyte effect on pH gradients in isoelectric focusing on polyacrylamide gels supports the hypothesis previously advanced by Murel et al., which relates pH gradient decay in electrofocusing to progressive protonation from the anodic pH gradient terminus.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A zwitterionic methacrylamide derivative, (3-sulfopropyl)dimethy1(3-methacrylamidopropyl) ammonium inner salt (MAPS), was synthesized and then copolymerized with N,N′-methylenebisacrylamide (Bis) to make gels carrying covalently bound sulfobetaine groups. Gels were also prepared with the addition of (3-sulfopropyl)trimethyl ammonium inner salt (TMAPS) to Bis-crosslinked polyacrylamide prior to polymerization. The properties of these gels in gel electrophoresis and electrofocusing were tested and compared with those of Bis-crosslinked polyacrylamide.In MAPS gels the absolute migration velocities of proteins and dyes at either polarity of migration were decreased. Anionic species were much more strongly retarded than cationic species. Ferguson plots (based on protein mobilities relative to dye in a continuous buffer) of catalase at pH 4 obtained on MAPS gels had a similar slope but lower y-intercept compared with those on equivalent polyacrylamide gels, indicating that the available (effective) net charge on the protein was decreased because it interacts with the zwitterionic charge on the gel. Similarly, the addition of TMAPS to polyacrylamide decreased the relative catalase mobility at non-restrictive gel concentrations. However, at gel concentrations above 4 %T, relative mobilities were increased, presumably because the effective pore size of the gel was increased through inhibition of polymerization by the zwitterionic compound.MAPS gels did not exhibit electroendosmosis by the criterion of cyanocobalamin displacement. Resolution between peptides in MAPS gels, and between proteins in ternary copolymer gels made of MAPS, Bis and polyacrylamide, was similar to that on polyacrylamide. Bands of catalase appeared sharpened in MAPS gels. pH Gradients form, decay and exhibit conductance gaps in MAPS electrofocusing gels and in polyacrylamide gels in presence of 0.25-1.5 M TMAPS, in a manner which was qualitatively indistinguishable from polyacrylamide.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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