Stable storage conditions of Immobiline chemicals for isoelectric focusing.
Identifieur interne : 002839 ( Ncbi/Curation ); précédent : 002838; suivant : 002840Stable storage conditions of Immobiline chemicals for isoelectric focusing.
Auteurs : B M G Veby [Suède] ; P. Pettersson ; J. Andrasko ; L. Ineva-Flygare ; U. Johannesson ; A. Görg ; W. Postel ; A. Domscheit ; P L Mauri ; P. PiettaSource :
- Journal of biochemical and biophysical methods [ 0165-022X ]
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Abstract
Most of the problems connected with the use of the Immobiline chemicals (a set of six, non-amphoteric, acrylamido buffers having pK values in the pH 3.5-9.5 interval) can be attributed to the alkaline species (with pK values 6.2, 7.0, 8.5 and 9.3). These compounds, to varying degrees are subjected to two degradation pathways: (a) hydrolysis of the amido bond, producing free acrylic acid and a diamine, the latter unable to be incorporated into the polyacrylamide matrix; (b) spontaneous auto-polymerization, producing a number of oligomers up to n-mers, able to aggregate and precipitate large proteins. Storage of their water solutions as frozen aliquots, a method widely employed, only partially alleviates the problem. Addition of trace-amounts of inhibitors, as lately adopted by the manufacturer, could only reduce the problem of auto-polymerization, but not block the hydrolysis of the amido bond. A new solution has been found, which abolishes both phenomena: storage in n-propanol. As demonstrated by gas chromatography, HPLC analyses and two-dimensional separations of complex samples, storage in organic solvent completely abolishes both hydrolysis and auto-polymerization and allows production of highly reproducible focusing patterns.
DOI: 10.1016/0165-022x(88)90026-7
PubMed: 3411080
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Ineva-Flygare</name></author><author><name sortKey="Johannesson, U" sort="Johannesson, U" uniqKey="Johannesson U" first="U" last="Johannesson">U. Johannesson</name></author><author><name sortKey="Gorg, A" sort="Gorg, A" uniqKey="Gorg A" first="A" last="Görg">A. Görg</name></author><author><name sortKey="Postel, W" sort="Postel, W" uniqKey="Postel W" first="W" last="Postel">W. Postel</name></author><author><name sortKey="Domscheit, A" sort="Domscheit, A" uniqKey="Domscheit A" first="A" last="Domscheit">A. Domscheit</name></author><author><name sortKey="Mauri, P L" sort="Mauri, P L" uniqKey="Mauri P" first="P L" last="Mauri">P L Mauri</name></author><author><name sortKey="Pietta, P" sort="Pietta, P" uniqKey="Pietta P" first="P" last="Pietta">P. Pietta</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="????"><PubDate><MedlineDate>1988 Jun-Jul</MedlineDate></PubDate></date><idno type="RBID">pubmed:3411080</idno><idno type="pmid">3411080</idno><idno type="doi">10.1016/0165-022x(88)90026-7</idno><idno type="wicri:Area/PubMed/Corpus">002A59</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002A59</idno><idno type="wicri:Area/PubMed/Curation">002A59</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002A59</idno><idno type="wicri:Area/PubMed/Checkpoint">002B22</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002B22</idno><idno type="wicri:Area/Ncbi/Merge">002839</idno><idno type="wicri:Area/Ncbi/Curation">002839</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Stable storage conditions of Immobiline chemicals for isoelectric focusing.</title><author><name sortKey="G Veby, B M" sort="G Veby, B M" uniqKey="G Veby B" first="B M" last="G Veby">B M G Veby</name><affiliation wicri:level="1"><nlm:affiliation>Pharmacia LKB Biotechnology AB, Bromma, Sweden.</nlm:affiliation><country xml:lang="fr">Suède</country><wicri:regionArea>Pharmacia LKB Biotechnology AB, Bromma</wicri:regionArea><wicri:noRegion>Bromma</wicri:noRegion></affiliation></author><author><name sortKey="Pettersson, P" sort="Pettersson, P" uniqKey="Pettersson P" first="P" last="Pettersson">P. Pettersson</name></author><author><name sortKey="Andrasko, J" sort="Andrasko, J" uniqKey="Andrasko J" first="J" last="Andrasko">J. Andrasko</name></author><author><name sortKey="Ineva Flygare, L" sort="Ineva Flygare, L" uniqKey="Ineva Flygare L" first="L" last="Ineva-Flygare">L. Ineva-Flygare</name></author><author><name sortKey="Johannesson, U" sort="Johannesson, U" uniqKey="Johannesson U" first="U" last="Johannesson">U. Johannesson</name></author><author><name sortKey="Gorg, A" sort="Gorg, A" uniqKey="Gorg A" first="A" last="Görg">A. Görg</name></author><author><name sortKey="Postel, W" sort="Postel, W" uniqKey="Postel W" first="W" last="Postel">W. Postel</name></author><author><name sortKey="Domscheit, A" sort="Domscheit, A" uniqKey="Domscheit A" first="A" last="Domscheit">A. 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These compounds, to varying degrees are subjected to two degradation pathways: (a) hydrolysis of the amido bond, producing free acrylic acid and a diamine, the latter unable to be incorporated into the polyacrylamide matrix; (b) spontaneous auto-polymerization, producing a number of oligomers up to n-mers, able to aggregate and precipitate large proteins. Storage of their water solutions as frozen aliquots, a method widely employed, only partially alleviates the problem. Addition of trace-amounts of inhibitors, as lately adopted by the manufacturer, could only reduce the problem of auto-polymerization, but not block the hydrolysis of the amido bond. A new solution has been found, which abolishes both phenomena: storage in n-propanol. As demonstrated by gas chromatography, HPLC analyses and two-dimensional separations of complex samples, storage in organic solvent completely abolishes both hydrolysis and auto-polymerization and allows production of highly reproducible focusing patterns.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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