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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 1 (1986), S. 43-46 
    ISSN: 0887-3585
    Keywords: protein stability ; helix-coil ; mutant ; calorimetry ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Oligonucleotide-directed mutagenesis has been used to replace α-helical glycines in the N-terminal domain of λ repressor with alanines. Since alanine is a significantly better helix-forming residue than glycine, these changes were predicted to have a stabilizing effect. We show that the Gly46→Ala substitution, the Gly48→Ala substitution, and the double substitution increase the melting temperature of the N-terminal domain by 3-6°.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 2 (1981), S. 320-323 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A study of properdin factor B (Bf) in 22 North and Central American populations demonstrated gene clusters delineating Old World Caucasian, New World and Black Ancestoral groups. Result of gene frequencies were comparable to data in the literature of European Caucasian and African Blacks. New World indigeneous population were clearly separated from other racial groups with hybrid population clustering in between the major genetic contributions.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Micro-polyacrylamide gel electrophoresis (micro-PAGE) has been shown to be useful for molecular size separation of urinary proteins. In addition to the sodium dodecyl sulfate micro-gradient gel electrophoresis in 10 μL capillaries, which is used routinely for examination of proteinuria in our laboratory, we tried micro-gradient slab gel electrophoresis (micro-slab-PAGE) for molecular weight determination of urinary proteins. It was also employed for monitoring patients with proteinuric diseases. For detailed studied of urinary proteins, micro-two-dimensional-electrophoresis is recommended.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 5 (1984), S. 133-138 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Two polyacrylamide gel electrophoretic systems for the separation of small peptides are described. In both systems the samples are concentrated by a buffer discontinuity, and the peptides separated according to size and charge in 40-50 % polyacrylamide resolving gels. The acidic gel has an operative pH between 3.0 and 3.1 while the alkaline gel has an operative pH between 9.1 and 9.2. The lower limit of resolution is about 100 daltons for a peptide with one positive charge in the acidic system. The radioactive peptides are detected by autoradiography. Peptides with a free amino group can be fixed in these gels with formaldehyde, so that fluorographic procedures can also be used for increased sensitivity. These techniques enable precise comparison of non-derivatized peptides from complete protein fingerprints and offer a simple one-dimensional analytical method for protein comparison, as well as a general method for separating and analyzing small peptides.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 0952-3499
    Keywords: monoclonal antibodies ; human ; antigen ; mouse ; allergen ; phospholipane ; epitope mapping ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Two human and twelve murine monoclonal antibodies directed against the main bee venom allergen phospholipase A2 (PLA) were evaluated for their fine specificity of binding to antigen and their ability to inhibit the enzymatic activity of the antigen. Antibodies were induced by natural exposure of beekeepers to bee venom or immunization of mice via different methods. Both human monoclonal antibodies (hmAbs) were previously shown to recognize the native three-dimensional conformation of PLA and are directed against discontinuous epitopes which include lysine residue at position 25 as a contact residue. In contrast, six of the murine monoclonal antibodies (mmAbs) bind to the denatured structure of the protein as determined by enzyme-linked immunosorbent assay. The epitopes recognized are located near the C-terminal end (n=8), in the centre of the polypeptide (n=1), near the N-terminal end (n=1) or include the carbohydrate part (n=2) of the PLA molecule. The capacity of the antibodies to modify the enzymatic activity was also determined. The hmAbs significantly inhibit the enzyme (70-79%), whereas the mmAbs produced various degrees of inhibition (39-100%). Since the X-ray structure of PLA is known, the epitopes can be visualized in the context of the three-dimensional structure of the antigen. A qualitative correlation was found between the location of epitopes and the inhibition pattern. Strong inhibition was seen with those antibodies that recognize epitopes that lie on the surface of the enzyme that is thought to contact the phospholipid bilayer. The results show that even though both hmAbs and most mmAbs inhibit the enzymatic activity of PLA, the antigen-binding properties of antibodies from different species raised after different routes of immunization differ significantly. Thus, detailed epitope mapping studies using murine antibodies prepared by artificial immunization may have limited value in predicting epitope patterns relevant to an antibody response to allergens in humans naturally exposed to antigen/allergen. © 1997 John Wiley & Sons, Ltd.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 32 (1988), S. 843-852 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The mechanism primarily implicated in the solventogenesis process in batch fermentations of Clostridium acetobutylicum is examined in considerable detail. A variety of fermentations with or without pH control in the pH range of 3.7-6 have been carried out in order to examine which of a host of suspect parameters correlate with the initiation of solventogenesis. The parameters that did not correlate are the external (pH0) and intracellular (pHi) pH, and ΔpH, and the external or intracellular butyrate and acetate concentrations. Undissociated butyric acid (UBA) correlated well with the initiation of solventogenesis. A linear relationship between UBA and butanol concentrations was found at the onset of solventogenesis in all fermentations examined. The intercept of this linear relationship was 6-13mM UBA for the pH0 range of 3.7-5 and approximately zero for pH0 at or above 6. The required minimal UBA was interpreted as a dependency of the solventogenesis process on both H+ and butyrate concentrations. Undissociated acetic acid was found not to correlate with the initiation of solventogenesis. Addition of acetoacetate (AA) and propionate enhanced the effect of UBA on the solventogenesis process. The action of a nonmetabolizable (FCCP) and a metabolizable (AA) uncoupler on the ΔpH, pH0, pHi, and solventogenesis were also studied in order to gain further understanding of the solventogenesis mechanism.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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