Journal article
Electrophoretic field gradient focusing: an investigation of the experimental parameters
Electrophoresis, Vol.29(2), pp.457-465
01/2008
Handle:
https://hdl.handle.net/2376/109845
PMID: 18064598
Abstract
Electrophoretic field gradient focusing has been used to separate the two oxidation states of myoglobin (Mb), and to separate Mb from bromophenol blue (BPB). Polyacrylamide and Sephadex were shown to be suitable packing materials whilst silica led to band broadening with Mb. BPB and Mb could be simultaneously focused apart using either a fixed 21-electrode setup or a dynamic 6-electrode setup. Using a dynamic three-electrode setup either analyte could be focused but not both simultaneously. It was shown that a higher ionic strength buffer in the separation channel compared to the coolant channel enhanced focusing between electrodes due to a conductivity gradient. Different running buffers were investigated and it was found that using a pH 8.6 buffer containing N,N,N-tris(hydroxymethyl)aminomethane (Tris) and phosphate ions the oxidation states of Mb could be separated but the separation of Mb from BPB was not as good as would be hoped for. Using a pH 8.6 buffer containing Tris, N-2-hydroxyethylpiperazine-N'-3-propanesulphonate and chloride ions as running buffer, BPB and Mb could be well separated but the two oxidation states of Mb merged.
Metrics
9 Record Views
Details
- Title
- Electrophoretic field gradient focusing: an investigation of the experimental parameters
- Creators
- Pilar González Tuñón - School of Chemistry, University of Leeds, Leeds, UKYating WangPeter MyersKeith D BartleLarry BowhillCornelius F IvoryRichard J Ansell
- Publication Details
- Electrophoresis, Vol.29(2), pp.457-465
- Academic Unit
- Chemical Engineering and Bioengineering, School of
- Publisher
- Germany
- Grant note
- BB/C006771/1 / Biotechnology and Biological Sciences Research Council
- Identifiers
- 99900547006001842
- Language
- English
- Resource Type
- Journal article