Polyacrylamide gel electrophoresis of soil humic acid fractionated by size-exclusion chromatography and ultrafiltration
A humic acid (HA) from chernozem soil has been fractionated either by size-exclusion chromatography (SEC) on Sephadex G-75 using water, Tris-HCl or 7 M urea as eluents, or by ultrafiltration (UF) in the presence of 7 M urea or water. UF-fractionated HA was classified as 100K (nominal retention size >100 000); 30K (100 000-30 000); 10K (30 000-10 000); 5K (10 000-5000). Several pools from each chromatography and all ultrafiltration retentates were assayed by polyacrylamide gel electrophoresis (PAGE). The results indicate that SEC fractionation gave a better separation of HA on fractions differing in electrophoretic mobility and molecular size (MS), and that apparently SEC is a more feasible technique than UF for soil HA fractionation; nevertheless the UF in 7 M urea solution might be useful for MS evaluation of HA fractions obtained by association SEC-PAGE. PAGE in the presence of denaturing agents can be successfully used for checking the purity of HA fractions obtained by both SEC and UF.
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Elsevier
1997
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dig-irnas-es-10261-586852018-09-13T09:13:10Z Polyacrylamide gel electrophoresis of soil humic acid fractionated by size-exclusion chromatography and ultrafiltration Trubetskoj, O. A. Trubetskaya, O. E. Afanas'eva, G. V. Reznikova, O. I. Sáiz-Jiménez, Cesáreo Junta de Andalucía Russian Academy of Sciences Consejo Superior de Investigaciones Científicas (España) A humic acid (HA) from chernozem soil has been fractionated either by size-exclusion chromatography (SEC) on Sephadex G-75 using water, Tris-HCl or 7 M urea as eluents, or by ultrafiltration (UF) in the presence of 7 M urea or water. UF-fractionated HA was classified as 100K (nominal retention size >100 000); 30K (100 000-30 000); 10K (30 000-10 000); 5K (10 000-5000). Several pools from each chromatography and all ultrafiltration retentates were assayed by polyacrylamide gel electrophoresis (PAGE). The results indicate that SEC fractionation gave a better separation of HA on fractions differing in electrophoretic mobility and molecular size (MS), and that apparently SEC is a more feasible technique than UF for soil HA fractionation; nevertheless the UF in 7 M urea solution might be useful for MS evaluation of HA fractions obtained by association SEC-PAGE. PAGE in the presence of denaturing agents can be successfully used for checking the purity of HA fractions obtained by both SEC and UF. The research described in this paper was made possible in part by grant MU000 from the Interna- tional Science Foundation (ISF) and cooperative grant M U 0 3 0 0 between the Russian Government and ISF. The authors also benefitted from the Cooperation Agreement between the Russian Academy of Sciences and the C.S.I.C. and a grant from the Junta de Andalucía for scientific exchanges. Peer Reviewed 2012-10-23T11:26:37Z 2012-10-23T11:26:37Z 1997 2012-10-23T11:26:37Z artículo http://purl.org/coar/resource_type/c_6501 doi: 10.1016/S0021-9673(97)00019-8 issn: 0021-9673 e-issn: 1873-3778 Journal of Chromatography A 767(1-2): 285- 292 (1997) http://hdl.handle.net/10261/58685 10.1016/S0021-9673(97)00019-8 http://dx.doi.org/10.13039/501100002674 http://dx.doi.org/10.13039/501100003339 http://dx.doi.org/10.13039/501100011011 en none Elsevier |
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A humic acid (HA) from chernozem soil has been fractionated either by size-exclusion chromatography (SEC) on Sephadex G-75 using water, Tris-HCl or 7 M urea as eluents, or by ultrafiltration (UF) in the presence of 7 M urea or water. UF-fractionated HA was classified as 100K (nominal retention size >100 000); 30K (100 000-30 000); 10K (30 000-10 000); 5K (10 000-5000). Several pools from each chromatography and all ultrafiltration retentates were assayed by polyacrylamide gel electrophoresis (PAGE). The results indicate that SEC fractionation gave a better separation of HA on fractions differing in electrophoretic mobility and molecular size (MS), and that apparently SEC is a more feasible technique than UF for soil HA fractionation; nevertheless the UF in 7 M urea solution might be useful for MS evaluation of HA fractions obtained by association SEC-PAGE. PAGE in the presence of denaturing agents can be successfully used for checking the purity of HA fractions obtained by both SEC and UF. |
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Junta de Andalucía |
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Junta de Andalucía Trubetskoj, O. A. Trubetskaya, O. E. Afanas'eva, G. V. Reznikova, O. I. Sáiz-Jiménez, Cesáreo |
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Trubetskoj, O. A. Trubetskaya, O. E. Afanas'eva, G. V. Reznikova, O. I. Sáiz-Jiménez, Cesáreo |
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Trubetskoj, O. A. Trubetskaya, O. E. Afanas'eva, G. V. Reznikova, O. I. Sáiz-Jiménez, Cesáreo Polyacrylamide gel electrophoresis of soil humic acid fractionated by size-exclusion chromatography and ultrafiltration |
author_sort |
Trubetskoj, O. A. |
title |
Polyacrylamide gel electrophoresis of soil humic acid fractionated by size-exclusion chromatography and ultrafiltration |
title_short |
Polyacrylamide gel electrophoresis of soil humic acid fractionated by size-exclusion chromatography and ultrafiltration |
title_full |
Polyacrylamide gel electrophoresis of soil humic acid fractionated by size-exclusion chromatography and ultrafiltration |
title_fullStr |
Polyacrylamide gel electrophoresis of soil humic acid fractionated by size-exclusion chromatography and ultrafiltration |
title_full_unstemmed |
Polyacrylamide gel electrophoresis of soil humic acid fractionated by size-exclusion chromatography and ultrafiltration |
title_sort |
polyacrylamide gel electrophoresis of soil humic acid fractionated by size-exclusion chromatography and ultrafiltration |
publisher |
Elsevier |
publishDate |
1997 |
url |
http://hdl.handle.net/10261/58685 http://dx.doi.org/10.13039/501100002674 http://dx.doi.org/10.13039/501100003339 http://dx.doi.org/10.13039/501100011011 |
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