Direct detection of carrier ampholytes in immobilized pH gradients using picric acid precipitation
| Autor: | L. Scott Rodkey, Keith A. Knisley |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
Acrylamides
Chromatography Resolution (mass spectrometry) Precipitation (chemistry) Isoelectric focusing Ampholyte Mixtures Clinical Biochemistry Proteins Picric acid Hydrogen-Ion Concentration Biochemistry Analytical Chemistry chemistry.chemical_compound Carrier Ampholytes Picrates chemistry Complex protein Chemical Precipitation Molecule Isoelectric Focusing |
| Zdroj: | Electrophoresis. 13:220-224 |
| ISSN: | 1522-2683 0173-0835 |
| DOI: | 10.1002/elps.1150130145 |
| Popis: | A protocol is described for monitoring the heterogeneity of end products of organic syntheses yielding amphoteric molecules containing two or more amino groups. This protocol was found to be a valuable aid in synthesis of carrier ampholytes for specific isoelectric focusing applications. This method does not depend on the ampholytes themselves to dictate the conditions under which they are analyzed. Carrier ampholytes have been found previously to be insoluble in picric acid and the insolubility property was not dependent upon the pI of individual ampholyte species. This insolubility property was exploited in the protocol. Immobilized pH gradients were used to focus the carrier ampholytes. Ampholytes were then visualized in situ by picric acid precipitation. The data shows that the protocol is useful for analyzing the results of chemical manipulations for enhancing the resolution of carrier ampholytes. A direct relationship was shown between carrier ampholyte heterogeneity as demonstrated by this protocol and the resolution of complex protein mixtures in isoelectric focusing gels. Picric acid formed visible precipitates with a variety of organic compounds which contained more than one amino group. |
| Databáze: | OpenAIRE |
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