Preparation of polyacrylamide gel‐filled capillaries for capillary electrophoresis

Dolnı́k, Vladislav; Cobb, Kelly A.; Novotný, Miloš V.

doi:10.1002/mcs.1220030209

Cited by 46 publications

(15 citation statements)

References 12 publications

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“…SDS PAGE has been adapted into capillary format with a crosslinked polyacrylamide gel as a sieving matrix [25,26], later the crosslinked gel was replaced with sieving polymer solutions [27–43] and the method was also modified for separations on microchip [44,45]. In capillary SDS electrophoresis, the anomalous migration of some proteins has been known for years.…”

Section: Introductionmentioning

confidence: 99%

Size separation of proteins by capillary zone electrophoresis with cationic hitchhiking

Dolnı́k¹,

Gurske²

2011

Electrophoresis

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The paper describes a method of size separation of proteins by capillary sieving electrophoresis with cationic surfactant. Proteins are separated within 12 minutes with repeatability of migration times better than 0.2%. Some proteins achieve the separation efficiency of 200,000 theoretical plates. The method can be used for determination of protein relative molecular masses. The accuracy of the determined relative molecular masses and the limitation of the method were investigated by the analysis of more than 60 proteins. The method also allows separation of protein oligomers. Proteins can be quantitated after the electrokinetic injection in the concentration range 0.07–0.43 g/L. The average detection limit is about 2 mg/L.

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Section: Introductionmentioning

confidence: 99%

Size separation of proteins by capillary zone electrophoresis with cationic hitchhiking

Dolnı́k¹,

Gurske²

2011

Electrophoresis

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“…However, gelfilled capillaries have several disadvantages: First, filling the capillary has to be done with great caution in order to avoid the introduction of air bubbles. The shrinkage of the gel during polymerization can also be a source of bubbles [4]. Gels, and in particular polyacrylamide, can be degraded by hydrolysis, especially at the alkaline pH normally used to separate biopolymers.…”

Section: Introductionmentioning

confidence: 99%

Finding a universal low‐viscosity polymer for DNA separation

Heller

1998

Electrophoresis

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We have investigated the viscosity of different commercially available polymers in solution and found that dextran has a low viscosity compared to other polymers of comparable molecular weight and resolving power. This makes it a potentially useful matrix for DNA separation in capillary electrophoresis, where either short time or low pressure are preferred for matrix replacement. We showed that dextran performs well for the separation of oligonucleotides and double-stranded DNA fragments. Together with the well-known application for protein separation, this makes dextran a universal polymer for the separation of biological macromolecules.

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“…Highly concentrated gels, immobilized in capillary tubes, 21 are suitable for the resolution of amino acid homopolymers, with the resolved peaks representing the individual oligomers. Thus, the charged Figure 1 (B) shows a unique splitting behavior not observed with the other polyamino acids.…”

Section: Results a N D Discussionmentioning

confidence: 99%

Electromigration behavior of poly‐(L‐glutamate) conformers in concentrated polyacrylamide gels

1993

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During electrophoretic separation of anionic polyamino acids, resolution according to the number of peptide units can be achieved in capillaries filled with hydrophilic gels. While polyaspartate preparations yield single peaks for the individual oligomers at pH above 8.0, polyglutamates exhibit an anomalous behavior of peak splitting, which is attributed here to the separation of the oligopeptide conformers. An Asp-Glu (1:1) copolymer yields single peaks under similar conditions. At pH near 4.5, where polyglutamate is expected to exist in its alpha-helix form, peak splitting disappears. Upon heating to 95 degrees C for at least 120 h (procedure described to transform the alpha-helix into a beta form), peak splitting disappeared, but could be reestablished after cooling for several days. When a highly charged cation spermine was added to the operational electrolyte, triple peaks appeared in the electropherogram due to the ion-pair formation. The largest peak in every triplet has been tentatively assigned to the alpha-helix form. The electrophoretic results described have been largely supported by CD spectra.

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Preparation of polyacrylamide gel‐filled capillaries for capillary electrophoresis

Cited by 46 publications

References 12 publications

Size separation of proteins by capillary zone electrophoresis with cationic hitchhiking

Size separation of proteins by capillary zone electrophoresis with cationic hitchhiking

Finding a universal low‐viscosity polymer for DNA separation

Electromigration behavior of poly‐(L‐glutamate) conformers in concentrated polyacrylamide gels

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