DD243565A1

DD243565A1 - ELECTROPHORESIS DEVICE FOR ISOELECTRIC FOCUSING IN GLASS TUBES

Info

Publication number: DD243565A1
Application number: DD28436485A
Authority: DD
Inventors: Hans Boernig
Original assignee: Univ Schiller Jena
Priority date: 1985-12-16
Filing date: 1985-12-16
Publication date: 1987-03-04

Abstract

Die Erfindung betrifft eine Elektrophoreseeinrichtung, die zur isoelektrischen Auftrennung von Proteinen und Biopolymeren in Glasroehrchen eingesetzt werden kann. Ziel und Aufgabe der Erfindung ist es, die isoelektrische Fokussierung in Glasroehrchen mit geringen Toleranzen in geringstmoeglichen Pufferloesungen durchzufuehren, die dabei auftretende Joulsche Waerme maximal abzufuehren und gleiche, reproduzierbare Wanderungsgeschwindigkeit in allen Roehrchen zu erreichen. Das Wesen der Erfindung besteht darin, dass fuer das untere Ende der Glasroehrchen jeweils eine mit einer Elektrode versehene Klammer zur Aufnahme der Pufferloesung vorgesehen ist, dass fuer das obere Ende der Glasroehrchen ebenfalls jeweils eine Elektrode angeordnet ist, die in den mit Puffer gefuellten Teil des Glasroehrchens hineinragt und dass Mittel zur Kuehlung der Glasroehrchen angebracht sind.The invention relates to an electrophoresis device which can be used for the isoelectric separation of proteins and biopolymers into glass tubes. The aim and object of the invention is to carry out the isoelectric focusing in glass tubes with small tolerances in the lowest possible buffer solutions, to minimize the occurring Joule heat and to achieve the same, reproducible migration speed in all cylinders. The essence of the invention is that for the lower end of the glass tubes each provided with an electrode clamp for receiving the buffer solution is provided, that for the upper end of the glass tubes also each an electrode is arranged in the filled with buffer part of the Glasroehrchens protrudes and that means for cooling the glass tubes are attached.

Description

embodiment

Ein Ausführungsbeispiel der Erfindung ist in der Zeichnung schematisch dargestellt und wird anschließend erläutert. Es zeigenAn embodiment of the invention is shown schematically in the drawing and will be explained. Show it

Fig. 1: die erfindungsgemäße Elektrophoreseeinrichtung im LängsschnittFig. 1: the electrophoresis device according to the invention in longitudinal section

Fig.2: das Oberteil der erfindungsgemäßen Elektrophoreseeinrichtung mit Elektroden in Aufsicht2: the upper part of the electrophoresis device according to the invention with electrodes in supervision

Fig.3: das Unterteil der erfindungsgemäßen Elektrophoreseeinrichtungen mit Kammern zur Aufnahme der Pufferlösung mit Elektroden in Aufsicht3 shows the lower part of the electrophoresis devices according to the invention with chambers for receiving the buffer solution with electrodes in plan view

Gemäß Fig. 1 trägt das Unterteil 1 vier Kammern zur Aufnahme der Pufferlösung 2 mit isolierten Elektroden 3 und vier Haltestützen 4 für das Mittelteil 5, eine davon dient zur Aufnahme der Steckbuchse 15. Im Mittelteil 5 sind Bohrungen 6 zur Halterung der Glasröhrchen 7 angebracht. Sie werden nach innen und außen mittels bekannter Gummiringe 8 abgedichtet. Eine verschließbare Öffnung 9 an der Oberseite dient zur Füllung des Mittelteiles 5 mit Kühlflüssigkeit. Zentral angeordnet ist eine Kühlschlange 10 mit Zu- und Abfluß, die über ein Kühlaggregat angeschlossen eine bedarfsgerechte Kühlung ermöglicht. Vier Stutzen 11 tragen das Oberteil 12 mit den Elektroden 3 und der Steckbuchse 15.1, the lower part 1 carries four chambers for receiving the buffer solution 2 with insulated electrodes 3 and four support posts 4 for the middle part 5, one of which serves to receive the socket 15. In the middle part 5 bores 6 for mounting the glass tube 7 are attached. They are sealed inwards and outwards by means of known rubber rings 8. A closable opening 9 on the upper side serves to fill the middle part 5 with cooling liquid. Centrally located is a cooling coil 10 with inflow and outflow, which connected via a cooling unit allows a needs-based cooling. Four sockets 11 carry the upper part 12 with the electrodes 3 and the socket 15.

Gemäß Fig. 2 wird das Aufsetzen des Oberteils 12 in seitenrichtiger Lage durch die Führungsnase 13 gewährleistet, die in eine entsprechende Nut des Halterungssteges auf dem Mittelteil 5 einrastet. Die Elektroden 3 sind so angeordnet, daß sie zentriert in die Glasröhrchen 7 eintauchen. Der Anschluß an das Stromversorgungsgerät erfolgt über die Steckbuchse 15. Gemäß Fig. 3 dient das Unterteil 1 zur Halterung von Mittelteil 5 und Oberteil 12 mittels der Haltestützen 4. Die seitenrichtige Lage wird durch die Nut 14 gewährleistet. Die Elektroden 3 sind seitlich an der Innenwand der Puffertröge 2 befestigt. Der Durchmesser der Puffertröge 2 ist dem der Glasröhrchen angepaßt. Die Steckbuchse 15 dient zum Anschluß an das Stromversorgungsgerät.According to Fig. 2, the placement of the upper part 12 is ensured in the correct position by the guide lug 13 which engages in a corresponding groove of the support web on the central part 5. The electrodes 3 are arranged so that they dive centered in the glass tube 7. The connection to the power supply device via the socket 15. According to FIG. 3, the lower part 1 serves to hold the middle part 5 and the upper part 12 by means of the support posts 4. The correct position is ensured by the groove 14. The electrodes 3 are attached laterally to the inner wall of the buffer troughs 2. The diameter of the buffer troughs 2 is adapted to the glass tube. The socket 15 is used for connection to the power supply unit.

Claims

Invention claim:

1. Elektrophoreseeinrichtung for isoelectric focusing in glass tubes, characterized in that for each of the lower end of the glass tube provided with an electrode chamber for receiving the buffer solution is provided, that for the upper end of the glass tube also in each case an electrode is arranged in immerse the buffered upper part of the glass tubes, and that the glass tubes are placed in a container having means for cooling them.

2. electrophoresis device according to item 1, characterized in that the chamber for receiving the buffer solution at the lower end of the glass tube to the diameter of the Glasröhrchen.angepaßt are.

For this

3 pages drawings

Field of application of the invention

The invention relates to a device that can be used for the isoelectric separation of proteins and biopolymers in glass tubes.

It finds application in biochemical and molecular biological basic research, biotechnology and medical diagnostics.

Characteristic of the known technical solutions

Electrophoresis devices in various designs are already known. For separations in glass tubes (disk electrophoresis), the devices consist of two separate buffer chambers and one electrode each, which are arranged one above the other. The carrier gel-coated glass tubes project into the bottom buffer chamber at one end and into the top buffer chamber at the other end. The sealing is done with O-rings. Disadvantageous for isoelectric focusing is the relatively large volume of the buffer chambers, which even with partial filling the risk of diffusion of sample material and buffer substance, as well as the different migration of the samples in the individual glass tubes, conditionally a) by the tolerance variations of the glass and b) by change the internal resistance of the Trägergele during the separation process and thus different temperature gradients in Gelro.hr. To eliminate these disadvantages, almost exclusively isoelectric focusing is carried out in a flat bed. However, these require a greater use of materials and increased consumption of chemicals and do not allow quantitative sample application.

Object of the invention

The aim of the invention is to enable a reliable time and cost-saving separation of macromolecular substances.

Explanation of the essence of the invention

The invention has for its object to carry out isoelectric focusing in glass tubes with low tolerances in the lowest possible buffer volume to dissipate the Joul heat occurring occurring maximum and to achieve the same, reproducible migration rates in all tubes. The object is achieved by means of an electrophoresis device for isoelectric focusing in glass tubes according to the invention in that for the lower end of the glass tube each provided with an electrode chamber for receiving the buffer solution is provided and the upper end of the glass tube above the gel contains a smaller buffer volume, in each case an electrode immersed directly, and that the glass tubes are guided in a container having means for cooling. In order to limit the amount of buffer and to minimize the loss of pH gradient substance, it is advantageous that the chambers for receiving the buffer solution at the lower end of the glass tube are adapted to the diameters of the glass tubes. By immersing the electrodes directly into the buffered upper end of the respective glass tubes, the diffusion of the applied samples is avoided. The inventive design of the electrophoresis is. ensures that the tubes are almost completely surrounded by coolant, thereby ensuring a uniform heat dissipation. The additional cooling device allows separations even under extreme, z. B / tropical conditions. The low buffer volume prevents loss of buffer material (to adjust the pH gradient), the adjustment of the pH gradient is relatively fast. In addition, the voltage drop is small. If the glass tubes are made of KPG glass and filled at the same time, the separation takes place reproducibly with the same migration speed.