JPH0196551A - Dispensing method of sample by electrophoresis - Google Patents

Abstract

PURPOSE:To take out fractionated components in a pure state as far as possible by separating a gel plate for migration in parallel with the development plane and using one thereof for specifying the components by dyeing and the other for collecting the components. CONSTITUTION:An isoelectric point migration soln. 4 is housed in a container 3 provided with electrodes 1, 2 at both ends and a potential gradient (pH gradient) is formed therein. A sample S is injected into such container. One end of the density gradient gel plate 5 consisting of acrylamide, etc., is then pressed to the migration liquid 4 to transfer the migrating components thereon. The fractionated components are further fractionate by the isoelectric point migration when electrodes 6, 7 are pressed to the side opposite to the transfer side and electric fields are applied thereto after the above-mentioned transfer. The gel plate 5 is, thereupon, separated in parallel with the development plane of the plate and one gel plate 5a is dyed by a silver dye, by which the development positions of the components are decided. The other gel plate 5b is floated on an electrolyte 8 provided with an electrode 9 and the other end of a capillary 11 provided with an electrode 10 at one end is pressed to the development position of the desired component. The desired component is moved by migration into the capillary if the electric field is impressed thereto. Only the desired component is obtd. by taking out the component from this capillary 11.

Description

DETAILED DESCRIPTION OF THE INVENTION (Technical Field) The present invention relates to a technique for fractionating a sample by electrophoresis and collecting target components.

(Prior art) When fractionating a substance consisting of many components, such as serum proteins, a first electrophoresis is usually performed by isoelectric focusing, and the fractionated product is transferred to an acrylamide gel plate. Precise fractionation is carried out by fractionating in the second direction by density gradient gel electrophoresis. By the way, when two-dimensionally fractionating in this way, the development position changes greatly depending on the electrophoresis conditions such as the component distribution of the electrophoresis gel plate and the electrophoresis time, making it unclear. Therefore, the gel plate is usually stained with silver and gold. Is the deployment position of the target component being determined? (Third point)
(Figure), there is a problem in that the binding of metals and proteins causes deterioration of the components, making it impossible to reuse the desired components.

(Purpose) The present invention was made in view of these problems, and its purpose is to provide a sample using electrophoresis that allows the fractionated components to be extracted in as pure a state as possible. The purpose of this research is to propose a preparative separation method.

(Summary of the Invention) In other words, the present invention is characterized in that a gel plate for electrophoresis is divided into two parts parallel to the development surface, and one part is used for identifying components by staining (the other part is used for collecting components). That's the point.

(Example) The details of the present invention will be described below based on illustrated examples.

FIG. 1 is an explanatory diagram showing the process of the present invention, in which an isoelectric focusing solution 4 such as amphorin is placed in a container 3 containing electrodes 1.2 at both ends, and an electric field is applied between the two electrodes. A potential gradient (PH gradient) is formed in the direction (I). After completing such preparation, when the sample S is injected into one end of the container 3, the components in the sample S move to the position where the pH reaches the isoelectric point and stop (II).

When the first development is completed in this manner, one end of the density gradient gel plate 5 made of acrylamide or the like is pressed against the electrophoresis liquid 4 to transfer the electrophoresis components ([1]). When this transfer is completed, there are electrodes 6. on the transfer side and on the other side.
7 and applies an electric field (■), migration starts in a direction perpendicular to the direction of isopotential migration, and each component fractionated by isopotential migration is roughly fractionated.

In this way, when the second-dimensional development is completed, the acrylamide gel plate is divided into two parts using a knife or the like parallel to the development surface (Vl).

One of the divided gel plates 5a is stained with silver dye to determine the development position at of the component.

On the other hand, the other gel plate 5bV divided into two is floating in the electrolytic solution 8, a first electrode 9 is provided in the electrolytic solution 8, and a thin tube 11 provided with an electrode 1o at one end is placed at the position where the target component is deployed.
The other ends are brought into contact and an electric field is applied between the two electrodes 7.8.

Thereby, the target component moves into the capillary 11 by electrophoresis.

When the components are taken out from this thin tube 11, only the target components can be obtained without containing dyes or gel plate components.

In this example, collection from the - point has been described, but by bringing a plurality of thin tubes into contact with each development point, it is possible to collect a plurality of components at the same time.

In addition, in this example, the position M of the tubules is determined while checking the development position H of the stained gel plate, but as shown in Fig. 2, the stained gel plate was
! Copy 12 is obtained by cutting on the contact glass surface of the copying device, and holes 13, 13, 13, etc. are punched at the positions where the target components are developed on the pattern of K and -12,
Alternatively, apply copy 12 to another board to make a mask.

By placing this mask on the surface of the other divided gel plate and abutting the capillary tube at the hole position, -
The target component can be reliably obtained in each layer.

(Effects) As explained above, according to the present invention, the electrophoresis plate is divided into two parts parallel to the development surface, and one part is used for identifying the component by staining, and the other part is used for collecting the component. So,
While accurately determining the deployment position M, it is possible to collect the target component in the purest possible state.

[Brief explanation of the drawing]

FIG. 1 is an explanatory diagram showing one embodiment of the present invention, FIG. 2 is a perspective view showing one embodiment of the mask used in the present invention, and FIG.
The figure is an explanatory diagram showing an example of a migration pattern by two-dimensional migration. ], 3... Electrode 4... Isoelectric focusing solution 5... Density gradient gel plate

Claims (1)

[Claims] A step of dividing a gel plate on which a sample has been developed by electrophoresis into two pieces parallel to the development surface, a step of determining the development position of the component by staining one of the gel plates, and a step of determining the development position of the component by staining one of the gel plates. A method for separating samples by electrophoresis, which consists of the step of collecting components by electrophoresis by touching one end of a thin tube in a position corresponding to the deployed position.