Substitution of salt with a zwitterion, betaine, in native-PAGE, SDS-PAGE and affinity chromatography

Izyanti Ibrahim, Paul C. Engel

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Betaine is one of the compatible solutes produced by halophiles to evade the effect of osmosis and avoid desiccation. In order to minimise the effect of high salt on both denaturing and non-denaturing electrophoresis, salt was substituted with betaine. Furthermore, the native-PAGE conditions previously proposed was optimised to give better outcomes. A resolving gel with 10% acrylamide gave better resolution compared to 15% acrylamide used earlier. A different formazan dye, INT, gave a more contrast colour when stained afterwards with Coomassie blue staining. Incubation conditions at 37°C with slow stirring also gave a sharper activity and protein bands. Protein samples dialysed with betaine prior running an SDS-PAGE managed to avoid the effect of high salt on gels. In addition, preliminary test using betaine instead of salt in halophilic NADP+-GDH purification using dye-ligand affinity columns showed positive results.

Original languageEnglish
Pages (from-to)11-15
Number of pages5
JournalMalaysian Journal of Biochemistry and Molecular Biology
Volume19
Issue number1-2
Publication statusPublished - 1 Aug 2016

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Native Polyacrylamide Gel Electrophoresis
Affinity chromatography
Betaine
Affinity Chromatography
Polyacrylamide Gel Electrophoresis
Substitution reactions
Salts
Acrylamide
Coloring Agents
Gels
Formazans
Osmosis
Desiccation
Distillation columns
Electrophoresis
NADP
Purification
Proteins
Color
Staining and Labeling

Keywords

  • Activity staining
  • Betaine
  • Dye-ligand columns
  • Halophile
  • Native-PAGE
  • SDS-PAGE

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Substitution of salt with a zwitterion, betaine, in native-PAGE, SDS-PAGE and affinity chromatography. / Ibrahim, Izyanti; Engel, Paul C.

In: Malaysian Journal of Biochemistry and Molecular Biology, Vol. 19, No. 1-2, 01.08.2016, p. 11-15.

Research output: Contribution to journalArticle

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