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  • Articles  (4,462)
  • Biochemistry and Biotechnology  (4,462)
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  • 1
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 7 (1986), S. 54-55 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The interpretation of nucleic acid sequences on polyacrylamide gels is often made difficult due to “smiles”. We have devised a simple modification for commonly used electrophoresis systems to eliminate this problem.
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  • 2
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 7 (1986), S. 56-57 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Lateral streaking or bending in vertical one-dimensional slab gel electrophoresis can be completely avoided if a comb-gel made up in electrophoresis buffer is placed on top of the stacking gel or sample gel.
    Additional Material: 3 Ill.
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  • 3
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 7 (1986), S. 58-58 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    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: The level of genetic variation revealed by two-dimensional electrophoresis of proteins from seedlings of two wheat lines strongly depends on the technical procedures. Improvements in extraction and electrophoresis procedures relative to earlier experiments on the same material led to a significant increase in the genetic variation revealed: 15.2 % instead of 6.7 % of the spots were genetically variable. The improved procedure is based on (i) precipipation of proteins from wheat seedlings with trichloroacetic acid and acetone, (ii) solubilization of the proteins with a solution containing urea, potassium carbonate and sodium dodecyl sulfate, (iii) isoelectric focusing in an optimized pH gradient, obtained with a mixture of carrier ampholytes (Pharmalyte and Servalyt), and (iv) running elecrophoresis in the second dimension on gels with increased surface.
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  • 5
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 2 (1981), S. 220-228 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The temperature at which polyacrylamide gels are polymerized strongly affects the structure and physical properties of this matrix. The suggested temperature of 0-4 °C has been found to produce highly turbid, highly porous and unelastic gels. At temperature of 25 °C of higher, the gels became progressively transparent, less porous and more elastic. This phenomenon is strongly pronounced in N,N′-methylenebisacrylamide (Bis) gels and progressively decreases for N,N′-(1,2-dihydroxyethylene)bisacrylamide (DHEBA) and N,N′-bisacrylylcystamine (BAC) gels. It has been attributed to the formation of hydrogen bonds among cross-linker molecules, which are presumably stabilized by low temperature and progressively broken at higher temperatures. The most extensively H-bonded compounds seem to be Bis molecules, since they can form four H-bonds/molecule, followed by DHEBA (in which inter-molecular H-bonds have to compete with intra-molecular H-bonds) and finally by BAC, which is sparingly H-bonded at 2 °C, and fully uncomplexed at 30°C. It is suggested that polymerization at 0-4°C should be abandoned, since it leads to unhomogeneous and unreproducible pore size matrices. Since H-bonds in Bis molecules are fully disrupted only at 60°C (a temperature unsuitable for gel polymerization, since it produces short chains and unelastic matrices), it is also suggested that the use of Bis should be discontinued in favour of better cross-links, such as DHEBA. In all cases, best polymerization conditions are obtained at 25-30°C. For all three cross-linkers, homogeneous gels are obtained in presence of 8 M urea or in pure formamide as solvent, since both agents fully disrupt H-bonds.
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  • 6
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 2 (1981), S. 228-235 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: An ultrasensitive silver stain is described for the detection of proteins in 4-20 % w/v polyacrylamide gradient gels of 3mm thickness, Maximal sensitivity was achieved by substituting methylamine for ammonia in the diamine preparation and adopting a strategy of overstain followed by destain. The prediamine steps were performed at 60°C to conserve time, and the intensity of final background stain was reduced by altering the method of diamine preparation. From our experience, the technique described is more sensitive and reliable than previously published methods when applied to gels of this thickness, In any case, the sensitivity in our hands was at least equal to that of the newly described photochemical technique [7].
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  • 7
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The rate of pH gradient development and protein migration towards their isoelectric points(pI) were investigated for thin-layer polyacrylamide gel isoelectric focusing (PAGIF) using Pharmalyte 3-10, Ampholine 3.5-9.5 and Servalyt 2-11 as carrier ampholytes. Using volt-hours(Vh) as the controlling parameter, it was found that both pH gradient development and protein focusing proceeded very similarly with all three carrier ampholytes, Only two significant (but minor) differences between the carrier ampholytes were observed: (1) due to a faster voltage rise with Pharmalyte, the Vh needed for focusing are achieved faster than with the other two carrier ampholytes and (2) focusing of protein with alkaline pI's (〉 pH 8) is slightly slower with Servalyt (both in time and Vh). This is presumably explained by the low field strength at the alkaline end of the pH gradient with Servalyt. Also, it was found that pH gradient development was completely independent of applied voltage when Vh were used as controlling parameter.
    Additional Material: 6 Ill.
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  • 8
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 7 (1986), S. 107-113 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: In order to survey changes in accumulation of several hundred proteins during the naturally synchronous nuclear division cycle of Physarum polycephalum, we have developed methods for analyzing two-dimensional (2-D) gel electrophoretograms using the general image processing system developed by the Spatial Data Analysis Laboratory at Virginia Tech. In this paper we describe fast and accurate methods for removing non-homogeneous background intensity from a 2-D gel image, for resolving overlapping protein spots, and for estimating the total integrated intensity in a protein spot by Gaussian modeling.
    Additional Material: 2 Ill.
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  • 9
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A method is described which allows the preparation of retinae, laminae, opticallobes, protocerebrum and proboscis of single heads of Drosophila. The protein content of these tissues was examined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Coomassie Brilliant Blue R-250 and glycoprotein staining. A catalog of 39 major proteins or protein subunits was obtained. Their relative proportions were determined quantitatively for the different tissues from Drosophila wildtype strains AS and C-S as well as the mutant sevLY3.
    Additional Material: 6 Ill.
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  • 10
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Electrophoresis 7 (1986), S. 221-226 
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A compact device, capable of being clipped onto the end of a 3 mm thick analytical polyacrylamide slab gel, has been developed for the collection of resolved protein zones as they elute from the end of the gel. Its unique feature is that eluted proteins were collected between two polyacrylamide-embedded paper membranes and removed either by continuous or intermittent buffer elution. Because of this direct coupling to large gels identical to those used for analytical electrophoresis, protein zones were obtained with high resolution. The method was applicable to non-denaturing cathodic and anodic electrophoresis, as well as sodium dodecyl sulfate electrophoresis. Medium-scale quantities (up to 10 mg/cm2 cross-sectional area) of protein were fractionated in as little as 4 h with high recoveries.
    Additional Material: 5 Ill.
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