JP5510247B2 - Gel cassette for two-dimensional electrophoresis and method for producing the same - Google Patents

Description

  The present invention relates to a gel cassette for two-dimensional electrophoresis and a method for producing the same.

  Electrophoresis technology that separates biopolymers such as DNA, RNA, and proteins extracted from animal and plant tissues and cells in electrophoresis gels based on differences in size, properties, etc. is a very important technology in the life science field. It is. In particular, when separating protein molecules by electrophoresis, a combination of SDS-polyacrylamide gel electrophoresis based on the difference in the molecular weight of the protein molecule and isoelectric focusing based on the difference in the isoelectric point of the protein molecule. Two-dimensional electrophoresis is widely used.

  As described above, the two-dimensional electrophoresis method can separate protein molecules based on the isoelectric point and the molecular weight. Therefore, it is difficult for the SDS-polyacrylamide gel electrophoresis method, which is a separation method of only the molecular weight. It is also possible to compare the expression differences of proteins having different isoelectric points but the same molecular weight. Therefore, more information is obtained than SDS-polyacrylamide gel electrophoresis, which is particularly useful in proteomics research and the like. For example, it is possible to apply to pathological diagnosis by comparing the expression of protein in a tissue of a healthy person and a diseased patient and detecting the difference. Such research is widely conducted and is expected to be used not only for basic research but also in clinical settings in the future.

  The gel cassette for two-dimensional electrophoresis used in the two-dimensional electrophoresis method once formed a gel cassette housing by combining a glass substrate and a spacer, and sealed one end portion of the gel cassette housing. Then, the experimenter himself prepared by filling the prepared gel layer forming solution from the other end portion and polymerizing the filled solution to form a gel layer. Gel cassettes called precast gels, in which a gel layer is formed in a gel cassette housing, are commercially available and widely used. Known gel cassettes for two-dimensional electrophoresis include, for example, those disclosed in Patent Document 1 below.

JP 2007-64848 A

  When producing a gel cassette for two-dimensional electrophoresis, as a method for forming a gel layer in the gel cassette housing, first, a gel solution is used to form a separation gel layer for separating and developing sample components. The cassette housing is filled, water or isobutanol or the like is layered thereon, and the separated gel solution is polymerized to form a separated gel layer. Next, the previously layered water or isobutanol is removed, and the formed gel layer is overlaid with a concentrated gel solution for forming a concentrated gel layer, and the concentrated gel solution is polymerized. To form a concentrated gel layer. In addition to such a two-stage filling method, there is also a continuous filling method in which a concentrated gel solution is carefully layered and filled immediately after the separation gel solution is filled.

  However, there are problems in both gel solution filling methods of the two-stage type and the continuous type. In the two-stage filling method, the interface between the two may be disturbed by the way of pouring water or isobutanol to be layered on the unpolymerized separated gel solution that has just been filled. There is a need to do. On the other hand, in the continuous filling method, it is necessary to carefully fill the concentrated gel solution so as not to disturb the interface of the previously filled separated gel solution and to prevent the gel solutions from mixing with each other as much as possible. . Therefore, in any of the filling methods, there is a problem that it is difficult to form a gel layer easily and with high reproducibility, which is troublesome.

  The present invention has been made in view of the above problems, and its object is to perform two-dimensional electrophoresis that can easily form a separation gel layer and a concentrated gel layer in a gel cassette housing with good reproducibility. An object of the present invention is to provide a gel cassette for use and a method for producing the same.

In order to achieve the above object, a method for producing a gel cassette for two-dimensional electrophoresis according to the invention described in claim 1 is a two-dimensional electrolysis in which a separation gel layer and a concentrated gel layer are accommodated in a gel cassette housing. In the method for producing a gel cassette for electrophoresis, a gel cassette housing, a separated gel solution that forms a separated gel by polymerization, and a concentrated gel solution that forms a concentrated gel by polymerization are prepared. A first sealed means is applied to the body to fill the separated gel solution, thereby defining a first sealed space filled with the separated gel solution, and the separated gel filled in the first sealed space The solution is polymerized to form a separated gel layer, and a second sealing means is applied to the gel cassette housing to fill the concentrated gel solution, thereby forming a second sealed space filled with the concentrated gel solution. Defined, the 2 sealed space by polymerizing the stacking gel solution filled in to form a concentrated gel layer, the gel cassette housing is composed of a portion retrofitting attached to the body portion and the body portion, the first enclosed space Is defined by the main body part of the gel cassette housing and the first sealing means before the retrofitting part is attached to the main body part, and the second sealed space is formed on the main body part. After the retrofit portion is attached, the main body portion, the retrofit portion, and the second sealing means are defined .

The invention according to claim 2 is the method described above, wherein the main body portion of the gel cassette housing has a pair of wall portions extending in parallel to each other with a space between each other. A first laminar space is formed between the walls, and the first hermetic space is defined by sealing the first laminar space with the first sealing means. The first sealing means And a peelable adhesive seal that seals one end of the laminar space and a removable sealing member that seals the other end of the laminar space.

The invention according to claim 3 is the method described above, wherein in the method, a second laminar space is formed between the main body portion and the retrofitting portion of the gel cassette housing, and the second laminar space is The second sealed space is defined by sealing with a second sealing means, and the second sealing means is an opening formed by the retrofitting portion of the gel cassette casing or the gel cassette casing. It comprises a removable sealing member for sealing an opening formed by the main body part and the retrofitting part, or a peelable adhesive sealing seal.

Moreover, the gel cassette for two-dimensional electrophoresis which concerns on invention of Claim 4 was manufactured by the said method.

  Since the separation gel solution and the concentrated gel solution are polymerized in a state where the sealed space is filled, respectively, the interface of the gel solution can be always kept uniform, whereby the separation gel layer and the concentration are concentrated in the gel cassette housing. The gel layer can be easily formed with good reproducibility.

A and B are diagrams showing a first specific example of a gel cassette housing in the gel cassette for two-dimensional electrophoresis according to the present invention, and C is a diagram showing a gel cassette housing according to a modification of the specific example. is there. A and B are diagrams showing a second specific example of a gel cassette housing in the gel cassette for two-dimensional electrophoresis according to the present invention, and C is a diagram showing a gel cassette housing according to a modification of the specific example. is there. It is the figure which showed the specific example of the manufacturing process of the gel cassette for two-dimensional electrophoresis which concerns on this invention. It is the figure which showed the 1st specific example of the 2nd sealing means applied to the gel cassette housing | casing of FIG. It is the figure which showed the 2nd specific example of the 2nd sealing means applied to the gel cassette housing | casing of FIG. It is the figure which showed the 3rd specific example of the 2nd sealing means applied to the gel cassette housing | casing of FIG. It is the figure which showed the 4th specific example of the 2nd sealing means applied to the gel cassette housing | casing of FIG. It is the figure which showed the 1st specific example of the 2nd sealing means applied to the gel cassette housing | casing of FIG. It is the figure which showed the 2nd specific example of the 2nd sealing means applied to the gel cassette housing | casing of FIG. It is the figure which showed the 3rd specific example of the 2nd sealing means applied to the gel cassette housing | casing of FIG. It is the figure which showed the 4th specific example of the 2nd sealing means applied to the gel cassette housing | casing of FIG.

  Hereinafter, specific embodiments of the two-dimensional electrophoresis gel cassette and the method for producing the same according to the present invention will be described. The gel cassette for two-dimensional electrophoresis according to the present invention comprises a separation gel layer and a concentrated gel layer housed in a gel cassette housing, and the manufacture of the gel cassette for two-dimensional electrophoresis according to the present invention. The method includes the steps of preparing a gel cassette housing, a separated gel solution that forms a separated gel by polymerization, and a concentrated gel solution that forms a concentrated gel by polymerization.

  1 and 2 show specific examples of the gel cassette housing. As shown in FIGS. 1A and 1B, the gel cassette housing 10 includes a gel cassette lid 12, a gel cassette substrate 14, and a concentrated gel surface base material 16, which are made of glass or acrylic resin. And an appropriate transparent material such as a styrene copolymer resin. Of these, the gel cassette lid 12 and the gel cassette substrate 14 are integrated by bonding, welding, or the like to form the main body portion 18 of the gel cassette housing. On the other hand, the concentrated gel surface base material 16 forms a rear part of the gel cassette housing. Therefore, the gel cassette housing 10 is composed of a main body part 18 and a rear part 16 attached to the main body part 18. Composed. In the method for manufacturing a gel cassette for two-dimensional electrophoresis according to the present invention, a separation gel layer is formed in the gel cassette housing 10 before attaching the retrofit portion 16 to the main body portion 18, and then the main body portion 18 is formed. A retrofit portion 16 is attached, and then a concentrated gel layer is formed in the gel cassette housing 10.

  FIG. 1A shows the gel cassette lid 12, the gel cassette substrate 14, and the concentrated gel surface base material 16 in a state before they are assembled to each other. FIG. 1B shows the gel cassette housing 10 in a completed state in which the gel cassette lid 12 and the gel cassette substrate 14 are integrated, and the concentrated gel surface base material 16 is further attached. . As shown in FIG. 1B, the gel cassette housing 10 includes a cathode buffer tank 20 and an anode buffer tank 22, and these buffer tanks 20 and 22 are formed in the main body 18. The main body portion 18 further includes a pair of wall portions 24 and 26 extending in parallel with each other with a space between each other, and the first thin layer space S1 is formed between the pair of wall portions 24 and 26. Is formed. As will be described later, the first thin layered space S1 is a separation gel layer forming space in which a separated gel solution is filled to form a separated gel layer, and one end of the first thin layered space S1 is The anode buffer tank 22 communicates with the slit-shaped opening 28, and the other end communicates with the cathode buffer tank 20 through the slit-shaped opening 30. The retrofit portion 16 is mounted in the cathode buffer tank 22, and a second thin layer space S <b> 2 is formed between the main body portion 18 and the retrofit portion 16 at the bottom of the cathode buffer tank 22. The second thin layered space S2 is a concentrated gel layer forming space in which the concentrated gel solution is filled to form a concentrated gel layer, which will be described later.

  In the specific examples shown in FIGS. 1A and 1B, the concentrated gel surface base material 16 is composed of a pair of parallel base plates having an elongated cross-sectional triangle extending in parallel. Once installed in the cathode buffer vessel 20, a slit-like opening 32 is defined between the pair of substrates. A part of the surface of the concentrated gel layer formed in the second laminar space can be exposed through the slit-shaped opening 32, and the gel subjected to the first-dimensional electrophoresis is removed from the cathode buffer tank 20. If the gel and the concentrated gel layer are brought into contact with each other in the region of the slit-shaped opening 32, the second-dimensional electrophoresis can be performed.

  FIG. 1C shows a part of the gel cassette housing 10 corresponding to a modification of the gel cassette housing 10 shown in FIGS. 1A and 1B. The body 10 differs from that shown in FIGS. 1A and 1B only in the form of the concentrated gel surface base material 16-1, which is the back part. This concentrated gel surface base material 16-1 connected both ends of a pair of triangular plate-shaped base materials constituting the concentrated gel surface base material 16 of FIGS. 1A and 1B, respectively. It is formed as a frame-shaped member, whereby a slit-shaped opening 32-1 is defined in the center of the concentrated gel surface base material 16-1. The function of the slit-shaped opening 32-1 is not different from the function of the slit-shaped opening 32 defined by the concentrated gel surface base material 16 described above.

  The gel cassette housing 10 of the specific example shown in FIGS. 2A and 2B is provided at the bottom of the cathode buffer tank 20 of the gel cassette housing 10 shown in FIGS. While forming the protruding line 34 for defining the edge part of concentrated gel layer formation space, the concentrated gel surface base material 16-2 which is a retrofit part is made of a plate-shaped base material of a pair of triangular sections. One is omitted, and only one plate-like base material having a triangular section is used. By providing the ridges 34, the concentrated gel layer forming space at the bottom of the cathode buffer tank 20 is reduced as compared with the gel cassette housing 10 of FIGS. 1A and 1B. The required amount of concentrated gel solution is reduced, and the capacity of the cathode buffer 20 is increased. A slit-shaped opening 32-2 is defined between the concentrated gel surface base material 16-2 and the ridge 34, and the function of the slit-shaped opening 32-2 is the same as that of the slit-shaped opening 32 described above. There is no difference with function.

  FIG. 2C shows a part of the gel cassette housing 10 corresponding to a modification of the gel cassette housing 10 shown in FIGS. 2A and 2B. The body 10 differs from that shown in FIGS. 2A and 2B only in the form of the concentrated gel surface base material 16-3, which is the back part. This concentrated gel surface base material 16-3 has short bent portions at both ends of a plate-shaped base material having a triangular cross section in the concentrated gel surface base material 16-2 in FIGS. It is formed as an added U-shaped member, thereby facilitating attachment of the concentrated gel surface base material 16-3 into the cathode buffer tank 20. A slit-shaped opening 32-3 is defined between the concentrated gel surface base material 16-3 and the ridge 34, and the function of the slit-shaped opening 32-3 is the same as that of the slit-shaped opening 32 described above. There is no difference with function.

  Hereinafter, an embodiment of a method for producing a gel cassette for two-dimensional electrophoresis according to the present invention will be described. FIG. 3 is a diagram showing a specific example of the manufacturing process of the gel cassette for two-dimensional electrophoresis of the present invention. 3A shows the main body portion 18 of the gel cassette housing before the retrofitting portion 16 is attached, and the first thin film is interposed between the pair of wall portions 24 and 26 of the main body portion 18. A layered space S1 (separation gel layer forming space) is formed. The slit-shaped opening 28 (on the anode buffer tank 22 side) at one end of the first thin-layer space is sealed with a peelable adhesive seal 36.

  The material of the sealing seal 36 may be any material that does not inhibit the polymerization of acrylamide, which is the main component of the separation gel and the concentration gel, and for example, polypropylene, polyvinyl chloride, polyester, or the like can be used. In addition, the pressure-sensitive adhesive that forms the pressure-sensitive adhesive layer of the sealing seal 36 for bonding to the gel cassette housing is a known type of pressure-sensitive adhesive called general high-viscosity, or high-viscosity. The type of pressure-sensitive adhesive can be used. However, since the gel cassette in which the gel layer is formed is generally stored at 4 ° C., it is more preferable that the gel cassette has low temperature resistance so that the adhesive strength does not decrease at low temperatures.

  Next, as shown in FIG. 3B, the separation gel solution is filled into the first thin layered space from the slit-shaped opening 30 (on the cathode buffer tank 20 side) at the other end of the first thin layered space. To do. Subsequently, as shown in FIG. 3C, the sealing member 38 is fitted into the cathode buffer tank 20, and the slit-shaped opening 30 on the cathode buffer tank 20 side of the first thin-layer space S1 is formed in this manner. Sealing is performed with a sealing member 38. The sealing member 38 can be removed.

  The material of the sealing member 38 may be the same material as the gel cassette housing, and a material that does not inhibit the polymerization of acrylamide, which is the main component of the separation gel and the concentrated gel, is used. Further, the size of the sealing member 38 only needs to be at least large enough to cover the slit-shaped opening 30, but may be the same size as the cathode buffer tank 20 in order to improve sealing performance.

  Then, the gel cassette housing | casing 10 is left still and the separation gel solution with which it filled is superposed | polymerized and a separation gel layer is formed. In the procedure so far, the sealing seal 36 and the sealing member 38 are the first sealing means applied to the gel cassette housing 10. Then, by applying the first sealing means to the gel cassette housing 10 and filling the separated gel solution, a first sealed space filled with the separated gel solution is defined, and the first sealed space is formed. The separation gel solution filled in is polymerized to form a separation gel layer. Further, the first sealed space has a first laminar space S1 formed between a pair of wall portions of the main body portion 18 before the retrofit portion 16 is attached to the main body portion 18 of the gel cassette housing 10. By being sealed by the first sealing means 36 and 38, the main body 18 and the first sealing means 36 and 38 are defined.

  Next, as shown in FIG. 3D, the sealing member 38 is removed from the cathode buffer tank 20, and the bottom of the cathode buffer tank 20 is filled with the concentrated gel solution as shown in FIG. . Subsequently, as shown in FIG. 3 (F), the concentrated gel surface base material 16 is covered with the gel cassette so as to cover the surface of the concentrated gel solution while taking care that air bubbles are not mixed into the packed concentrated gel solution. It is attached to the main body 18 of the housing 10. Further, as shown in FIGS. 3G and 3H, a sealing member 40 is fitted into the cathode buffer tank 20, and the slit-like opening 32 described above is sealed by the sealing member 40. Stop.

  Then, the gel cassette housing | casing 10 is left still and the filled concentrated gel solution is superposed | polymerized and a concentrated gel layer is formed. In the procedure so far, the sealing member 40 is the second sealing means applied to the gel cassette housing 10. Then, by applying the second sealing means to the gel cassette housing 10 and filling the concentrated gel solution, a second sealed space filled with the concentrated gel solution is defined, and this second sealed space is defined. The concentrated gel solution filled in is polymerized to form a concentrated gel layer. The second sealed space is a second laminar space S <b> 2 formed between the main body 18 and the rear part 16 after the rear part 16 is attached to the main body 18 of the gel cassette housing 10. Is sealed by the second sealing means 40, and is defined by the main body portion 18, the retrofit portion 16, and the second sealing means 40.

  The sealing member 40 can be removed, and is removed when the gel cassette for two-dimensional electrophoresis is used. The material of the sealing member 40 can be various materials similar to the sealing member 38 described above.

  In the above procedure for forming the concentrated gel layer, as shown in FIGS. 3E and 3F, the concentrated gel solution is filled first, and then the concentrated gel surface base material which is the back part. However, if there is no risk of air bubbles being mixed into the solution when the concentrated gel solution is filled, the concentrated gel surface substrate 16 is first attached, It is good also as a procedure which fills a concentrated gel solution after that.

  FIG. 4 is an enlarged view of a part of FIG. 3H showing the sealing member 40 in detail. This enlarged view shows a cathode that is a concentrated gel layer forming region of a gel cassette for two-dimensional electrophoresis. The vicinity of the buffer tank 20 is shown. The sealing member 40 is a first specific example of the second sealing means applied to the gel cassette housing 10 of FIG. As shown in the figure, the sealing member 40 includes a knob 42, and when the gel cassette for two-dimensional electrophoresis is used, the knob 42 is picked and the sealing member 40 is removed for use. 4A shows the sealing member 40 before mounting, and FIG. 4B shows the sealing member 40 after mounting. The mounted sealing member 40 is the concentrated gel surface base material 16. Adhering to the top surface

  5, 6, and 7 are views showing second, third, and fourth specific examples of the second sealing means applied to the gel cassette housing 10 of FIG. 1, and are the same as FIG. 4. Shows the vicinity of the cathode buffer tank 20. The sealing member 40-1 shown in FIG. 5 is a removable sealing member that is larger than the sealing member 40 shown in FIG. 4 and engages with a wide area on the inner surface of the cathode buffer tank 20. And includes a knob 42-1. Due to this increase in size, the adhesion to the gel cassette housing 10 is improved and it is more difficult to come off, and the stability is excellent. In the case where sufficient adhesion can be obtained only by engaging the wall surfaces at both ends in the longitudinal direction of the cathode buffer tank 20 and the end surfaces at both ends in the longitudinal direction of the sealing member, the sealing member is thinned on the contrary. It is also possible to cover only the slit-shaped opening 32 described above.

  The specific examples shown in FIGS. 6 and 7 are provided with peelable adhesive sealing seals 44 and 44-1 similar to the sealing seal 36 of the first sealing means instead of the removable sealing member. Two sealing means are used. The sealing seals 44 and 44-1 are provided with knobs 46 and 46-1, and when the gel cassette for two-dimensional electrophoresis is used, the sealing seal 44 is picked up by picking the knobs 46 and 46-1. 44-1 can be easily peeled off. The sealing seal 44 shown in FIG. 6 is made to adhere not only to the concentrated gel surface base material 16 but also to a part of the inner surface of the cathode buffer tank 20, and has high adhesion. On the other hand, the sealing seal 44-1 of FIG. 7 is made to adhere only to the concentrated gel surface base material 16, and the sticking work of the sealing seal is easy. 6A shows the sealing seal 44 before mounting, and FIG. 6B shows the sealing seal 44 after mounting. The mounted sealing seal 44 is the concentrated gel surface base material 16. Adhering to the top surface

  8, 9, 10, and 11 are views showing first, second, third, and fourth specific examples of the second sealing means applied to the gel cassette housing 10 of FIG. Yes, the vicinity of the cathode buffer tank 20 is shown as in FIGS. The second sealing means in FIG. 8 and FIG. 9 includes removable sealing members 40-2 and 40-3 similar to the sealing members 40 and 40-1 in FIG. 4 and FIG. 2, 42-3. The sealing member 40-2 of FIG. 8 covers the area | region from the concentrated gel surface base material 16-2 to the protruding item | line 34 among the bottom surfaces of the cathode buffer tank 20. FIG. On the other hand, the sealing member 40-3 in FIG. 9 covers the entire bottom surface of the cathode buffer tank 20. 8A shows the sealing member 40-2 before mounting, and FIG. 8B shows the sealing member 40-2 after mounting. The mounted sealing member 40-2 is shown in FIG. The concentrated gel surface substrate 16-2 is in close contact with the upper surface.

  The specific examples shown in FIGS. 10 and 11 use the adhesive sealing seals 44-2 and 44-3 that can be peeled off as the second sealing means in the same manner as the specific examples in FIGS. . These sealing seals 44-2 and 44-3 are provided with knobs 46-2 and 46-3. When this two-dimensional electrophoresis gel cassette is used, the knobs 46-2 and 46-3 are clamped. Thus, the sealing seals 44-2 and 44-3 can be easily peeled off. The sealing seal 44-2 in FIG. 10 is made to adhere to a part of the inner surface in addition to substantially the entire bottom surface of the cathode buffer tank 20, and has high adhesion. On the other hand, the sealing seal 44-3 in FIG. 11 covers only the region from the concentrated gel surface base material 16-2 to the ridges 34 and adheres only to them. Is easy. FIG. 10A shows the sealing seal 44-2 before mounting, and FIG. 10B shows the sealing seal 44-2 after mounting. The concentrated gel surface substrate 16-2 is in close contact with the upper surface.

  Thus, by using the method for producing a gel cassette for two-dimensional electrophoresis according to the present invention, it is possible to form the concentrated gel layer and the separated gel layer uniformly with good reproducibility. It is not necessary to overlay water or isobutanol after filling the separated gel solution as in the conventional gel cassette manufacturing method described above. In addition, since the operation of continuously filling the separated gel solution and the concentrated gel solution is not performed, it is not necessary to perform the filling operation so as not to disturb the interface of the gel solution, and the gel solution can be filled easily. It becomes possible.

  The present invention is not limited to the above-described embodiments, and various modifications can be made within the scope shown in the claims. In other words, embodiments obtained by combining technical means appropriately modified within the scope of the claims are also included in the technical scope of the present invention.

  By using the present invention, it becomes possible to provide a gel cassette for two-dimensional electrophoresis with good reproducibility, especially in two-dimensional electrophoresis of proteins such as proteomics, and thus an electrophoretic image with excellent reproducibility. Can be obtained. In addition, since the gel solution can be filled more easily and inferior to the conventional gel cassette, mass production is also easy. Furthermore, by further developing research on biopolymers such as DNA, RNA, and proteins, it can contribute to the development of industries in the fields of medicine, biology, and chemistry.

10 Gel cassette housing 12 Gel cassette lid 14 Gel cassette substrate 16 Concentrated gel surface base material (rear part of gel cassette housing)
18 Main Body of Gel Cassette Housing 20 Cathode Buffer Tank 22 Anode Buffer Tank 24 (Gel Cassette Cover) Wall 26 (Gel Cassette Substrate) Wall 28 Slit Open 30 Slit Open 32 Slit Open 34 Projection 36 Sealing Seal (first sealing means)
38 Sealing member (first sealing means)
40 sealing member (second sealing means)
42 Picking part 44 Sealing seal (second sealing means)
46 (seal) knob S1 first thin layered space S2 second thin layered space

Claims (4)

In the method for producing a gel cassette for two-dimensional electrophoresis in which a separated gel layer and a concentrated gel layer are housed in a gel cassette housing,
Prepare a gel cassette housing, a separated gel solution that forms a separated gel by polymerization, and a concentrated gel solution that forms a concentrated gel by polymerization,
By applying a first sealing means to the gel cassette housing and filling the separated gel solution, a first sealed space filled with the separated gel solution is defined, and the first sealed space is filled. The separated gel solution is polymerized to form a separated gel layer,
By applying a second sealing means to the gel cassette housing and filling the concentrated gel solution, a second sealed space filled with the concentrated gel solution is defined, and the second sealed space is filled. The concentrated gel solution is polymerized to form a concentrated gel layer ,
The gel cassette housing is composed of a main body part and a retrofit part attached to the main body part,
The first sealed space is defined by the main body portion of the gel cassette housing and the first sealing means before the retrofit portion is attached to the main body portion,
The second sealed space is defined by the main body part, the rear part, and the second sealing means after the rear part is attached to the main body part.
A method for producing a gel cassette for two-dimensional electrophoresis, characterized in that The body portion of the gel cassette housing has a pair of wall portions extending in parallel with each other with a space between each other, and a first laminar space is formed between the pair of wall portions, The first sealed space is defined by sealing the first laminar space with the first sealing means,
The first sealing means includes a peelable adhesive sealing seal that seals one end of the laminar space, and a removable sealing member that seals the other end of the laminar space.
The method for producing a gel cassette for two-dimensional electrophoresis according to claim 1 . A second laminar space is formed between the body portion and the retrofitting portion of the gel cassette housing, and the second hermetic seal is formed by sealing the second laminar space with the second sealing means. A space is defined,
The second sealing means is removable to seal the opening formed by the retrofit portion of the gel cassette housing or the opening formed by the main body portion and the retrofit portion of the gel cassette housing. Consisting of a sealing member or a peelable adhesive sealing seal,
The method for producing a gel cassette for two-dimensional electrophoresis according to claim 2 . A gel cassette for two-dimensional electrophoresis manufactured by the method for manufacturing a gel cassette for two-dimensional electrophoresis according to any one of claims 1 to 3 .

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