Micropreparative isoelectric focusing protein separation in a suspended drop

Egatz-Gomez, Ana; Thormann, Wolfgang (2011). Micropreparative isoelectric focusing protein separation in a suspended drop. Electrophoresis, 32(12), pp. 1433-7. Weinheim: Wiley-VCH 10.1002/elps.201000684

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IEF protein binary separations were performed in a 12-μL drop suspended between two palladium electrodes, using pH gradients created by electrolysis of simple buffers at low voltages (1.5-5 V). The dynamics of pH gradient formation and protein separation were investigated by computer simulation and experimentally via digital video microscope imaging in the presence and absence of pH indicator solution. Albumin, ferritin, myoglobin, and cytochrome c were used as model proteins. A drop containing 2.4 μg of each protein was applied, electrophoresed, and allowed to evaporate until it splits to produce two fractions that were recovered by rinsing the electrodes with a few microliters of buffer. Analysis by gel electrophoresis revealed that anode and cathode fractions were depleted from high pI and low pI proteins, respectively, whereas proteins with intermediate pI values were recovered in both fractions. Comparable data were obtained with diluted bovine serum that was fortified with myoglobin and cytochrome c.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases > Laboratory for Clinical Pharmacology

UniBE Contributor:

Thormann, Wolfgang

ISSN:

0173-0835

Publisher:

Wiley-VCH

Language:

English

Submitter:

Factscience Import

Date Deposited:

04 Oct 2013 14:25

Last Modified:

05 Dec 2022 14:07

Publisher DOI:

10.1002/elps.201000684

PubMed ID:

21626519

Web of Science ID:

000292256100004

URI:

https://boris.unibe.ch/id/eprint/8911 (FactScience: 214563)

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