Comparison of different techniques for isoelectric focusing on polyacrylamide gel slabs using bacterial asparaginases
| Autor: | H.K. Robinson |
|---|---|
| Rok vydání: | 1972 |
| Předmět: |
Electrophoresis
Asparaginase Time Factors Resolution (mass spectrometry) Polymers Biophysics Analytical chemistry Biochemistry chemistry.chemical_compound Escherichia coli Methods Molecular Biology Polyacrylamide gel electrophoresis chemistry.chemical_classification Acrylamides Chromatography Chemistry Isoelectric focusing Temperature Cell Biology Polymer Hydrogen-Ion Concentration Isoelectric point Polymerization Erwinia Electrophoresis Polyacrylamide Gel Isoelectric Focusing Gels |
| Zdroj: | Analytical Biochemistry. 49:353-366 |
| ISSN: | 0003-2697 |
| Popis: | The possibility has been investigated with bacterial asparaginases that the multiple bands from electrophoretically homogeneous proteins in polyacrylamide gel isoelectric focusing may be due to artifacts or experimental variation. Although the pH profile, and hence the resolution, in different areas can be altered by washing the polymerized gel, the type of polymerization used, and the application of the carrier ampholytes, the isoelectric points of the individual asparaginase preparations are similar. An improved resolution in the basic region of the gel can be obtained by employing a refrigerated platen in contact with the glass base-plate. Varying the position of the sample application does not alter the isoelectric points in a standard 24 hr run on a narrow-range gel with the acidic asparaginase preparation, whereas the basic asparaginases require a longer period (36 hr) to reach equilibrium. When excised focused protein bands are rerun, identical isoelectric points are obtained. The formation of artifacts from protein-ampholyte buffer interaction is therefore ruled out. |
| Databáze: | OpenAIRE |
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