JPH0674935A - Acrylic amide gel preserving method - Google Patents

Abstract

PURPOSE:To provide a highly reliable preserving method for acrylic amide gel used as a support body in electrophoresis. CONSTITUTION:In an electrophoresis device formed by using acrylic amide gel as a support body, after electrophoresis is carried out on a specimen by impressing a direct current electric field, a gel cover 5 having grater shape fine projections 6 is brought into contact with the surface of the acrylic amide gel sticking on a glass plate or a plastic plate as a method to preserve the specimen which is dyed by using pigment and to which a concentration difference is applied. Thereby, an acrylic amide gel preserving method can be constituted.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a method for storing an acrylamide gel used as a support for electrophoresis in an electrophoresis apparatus.

When a direct current voltage is applied to a molecule such as a protein existing in a liquid, + or-depending on the nature or shape of the molecule.
Molecules having a positive charge and a positive charge are directed toward the cathode,
In addition, there is a phenomenon that molecules having a negative charge move toward the anode.

This phenomenon is called electrophoresis, and here, the charged state of the molecule is related to the pH of the electric field. For example, when a protein exists in an acidic environment, it has a positive charge and goes toward the cathode. Also, when it exists in an alkaline environment, it has a negative charge and moves toward the anode.

Such electrophoresis is an indispensable technique for measuring the molecular weight, determining the base sequence, etc., including the separation of proteins, and is widely used in the research dealing with biopolymers.

Here, holding the solution containing the biopolymer,
Acrylamide gel, cellulose acetate, etc. are used as a support to apply a DC electric field to this, but the region (fraction) separated by electrophoresis is clear,
It is necessary to be sensitive and have excellent reproducibility of fractional values, and therefore acrylamide gel is often used.

[0006]

2. Description of the Related Art Methods for analyzing proteins by electrophoresis include SDS-PAGE method (sodium dodecyl sulfate-polyacrylamide gel electrophoresis method), Native-PAGE method and isoelectric focusing method.

Conventionally, the size of the support is about 12
An acrylamide gel of about 0 × 120 × 1 mm is used, and after the electrophoretic treatment is completed, in order to save the result, the film-shaped acrylamide gel is peeled off from the gel plate made of glass or acrylic resin. Then, it was sandwiched between filter paper and cellophane paper, and dried by vacuum suction.

However, the technique for separating proteins has improved, and the development of high-sensitivity staining method has made it possible to perform analysis even when the size of the support is small. Therefore, at present, about 40 × 50 × 0.
Supports as small as 3 mm have come to be used.

FIG. 2 shows the shape of such a miniaturized support. A gel plate 1 made of acrylic resin has a support 2 made of acrylamide gel on which an electrophoretic pattern 3 is formed by dyeing. Shows the state in which is formed.

The upper protrusion is the handle 4. The thickness of the support 2 made of acrylamide gel is 0.3 m.
Since it is as thin as about m, it is difficult to separate it from the gel plate 1.

For this reason, the gel plate 1 was dried until it was stored. However, if the drying is insufficient, other gels, paper, etc. will adhere to the acrylamide gel and it will not be able to be removed.

Further, there is a problem that it becomes sticky when exposed to moisture, and other gels, papers, etc. adhere to it, making it difficult to store.

[0013]

Acrylamide gel is used as a support for electrophoresis of polymer analytes in an electrophoresis apparatus because of its excellent resolution.

However, since the acrylamide gel used as a support has a thin thickness of about 0.3 mm and is difficult to peel off from the gel plate, it is stored as it is attached to the gel plate. It has a strong hygroscopic property, it is not easy to completely dry it, and it becomes sticky when it absorbs moisture, making it difficult to handle.

Therefore, conventionally, a method of taking a photograph and then discarding the test sample has been adopted.

[0016]

[Means for Solving the Problems] The above-mentioned problems are solved by contacting a gel cover having fine protrusions in the form of a metal on an acrylamide gel attached as a support on a gel plate made of a glass plate or a plastic plate. This can be solved by configuring a method for storing an acrylamide gel, which is characterized in that

[0017]

[Function] A material such as acrylamide gel, which easily absorbs moisture, tends to adhere to paper or other gel, and once attached, cannot be removed.

Further, in order to completely dry it, it may be stored in a desiccator or the like, but the handling is complicated. In view of this, the present invention prevents the adhesion of other acrylamide gel, paper, or dust to the acrylamide gel by bringing the gel cover having the fine projections in the shape of a metal down into contact with the surface.

FIG. 1 shows an example of the gel cover 5.
Many protrusions 6 are arranged to minimize contact with the acrylamide gel. The constituent material of the protrusion 6 may be metal, plastic or ceramic.
It should be as thin and light as possible to facilitate handling.

When such a gel cover is used, it can be easily taken for re-inspection, and since the contact area is small, the result of electrophoresis is not disturbed. It is also possible to stack and store the samples.

[0021]

【Example】

Example 1 (corresponding to claim 2) An aluminum (Al) foil having a thickness of 150 μm was press-molded to make a gel cover having the shape shown in FIG.

Next, E. coli HB101 (pYEJ001) was used as a sample.
Chloramphenicol acetyltransferase
Rase abbreviated CAT using an electrophoresis device (named Phast System, manufactured by Pharmacia) and acrylamide gel (named Phastg
Electrophoresis was carried out on a support consisting of el 10-15% (Pharmacia), followed by staining with a dye (Phastgel Blue R, Pharmacia).

After drying the glass gel plate at room temperature for 3 days, the gel cover made of Al foil was contacted and stored. Next, after one week, the gel cover was removed and observed, but the acrylamide gel membrane was not damaged and was not contaminated. Example 2: (corresponding to claim 3) An acrylic resin was molded to prepare a gel cover having the shape of FIG.

Then, the same sample as in Example 1 was used for electrophoresis, and the same dye was used for staining. Then, it was dried at room temperature for 3 days together with a glass gel plate, and then stored by contacting it with a gel cover made of plastics.

Next, after one week, the gel cover was removed and observed, but the acrylamide gel film was not damaged and was not contaminated.

[0026]

EFFECTS OF THE INVENTION By carrying out the present invention, it is possible to store a support made of acrylamide gel which has been subjected to electrophoresis treatment without causing contamination or damage, thereby improving the analysis efficiency of biopolymers. it can.

[Brief description of drawings]

FIG. 1 is a perspective view showing an example of a gel cover.

FIG. 2 is a configuration diagram of a support used for electrophoresis.

[Explanation of symbols]

 1 gel plate 2 support 3 electrophoresis pattern 5 gel cover 6 protrusions

Claims (3)

[Claims] 1. An electrophoretic apparatus using acrylamide gel as a support, a sample is electrophoresed by applying a DC electric field, and then stained with a dye to store the sample with a different concentration. As a method, a method for preserving acrylamide gel, characterized in that a gel cover having fine protrusions in the form of a down metal is brought into contact with the acrylamide gel attached on a gel plate made of a glass plate or a plastic plate. .. 2. The method for storing acrylamide gel according to claim 1, wherein the gel cover is made of metal. 3. The method for preserving acrylamide gel according to claim 1, wherein the gel cover is made of plastics.