JPS61104247A - 2D electrophoresis device - Google Patents
2D electrophoresis deviceInfo
- Publication number
- JPS61104247A JPS61104247A JP59225740A JP22574084A JPS61104247A JP S61104247 A JPS61104247 A JP S61104247A JP 59225740 A JP59225740 A JP 59225740A JP 22574084 A JP22574084 A JP 22574084A JP S61104247 A JPS61104247 A JP S61104247A
- Authority
- JP
- Japan
- Prior art keywords
- agarose
- syringe
- dimensional
- moving
- dimensional electrophoresis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000000539 two dimensional gel electrophoresis Methods 0.000 title claims description 14
- 229920000936 Agarose Polymers 0.000 claims abstract description 24
- 238000001962 electrophoresis Methods 0.000 claims description 14
- 230000001143 conditioned effect Effects 0.000 abstract 1
- 239000000499 gel Substances 0.000 description 16
- 238000000034 method Methods 0.000 description 7
- 229920002401 polyacrylamide Polymers 0.000 description 6
- 239000000758 substrate Substances 0.000 description 5
- 239000011544 gradient gel Substances 0.000 description 3
- 230000005012 migration Effects 0.000 description 3
- 238000013508 migration Methods 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 239000011521 glass Substances 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229920000728 polyester Polymers 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000006087 Silane Coupling Agent Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 239000012774 insulation material Substances 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
【発明の詳細な説明】
〔発明の利用分野〕
本発明は主として蛋白質の多成分同時分析を可能にする
自動２次元電気泳動装置の構造に関する。DETAILED DESCRIPTION OF THE INVENTION [Field of Application of the Invention] The present invention primarily relates to the structure of an automatic two-dimensional electrophoresis apparatus that enables simultaneous multi-component analysis of proteins.
２次元電気泳動法は１次元泳動用支持体の１個所に試料
を充填して１回目の泳動分離を行なったのち、この支持
体を２次元泳動用支持体と密着させて通電し、−２次元
泳動像を得る。従来垂直式２次元電気泳動法ではＪ 、
Ｂ ｉｏｃｈｅｍ、 ８５６４９ （１９７９）にみ
られるような円柱状の１次元泳動用支持体（ポリアクリ
ルアミドゲル、５ｍｎφＸ１４０ｎｗｎ）を２次元泳動
用支持体（濃度勾配ポリアクリルアミドゲル、１６０Ｘ
１４０Ｘ４ｍｍ、４〜２１％）の上部にのせ通電する。In the two-dimensional electrophoresis method, a sample is filled in one part of a one-dimensional electrophoresis support and a first electrophoretic separation is performed, and then this support is brought into close contact with a two-dimensional electrophoresis support and an electric current is applied. Obtain a dimensional electrophoresis image. In the conventional vertical two-dimensional electrophoresis method, J,
Biochem, 85649 (1979), a cylindrical one-dimensional electrophoresis support (polyacrylamide gel, 5 mmφ
140x4mm, 4-21%) and energize it.
このほかＪ 、 Ｂ ｉｏＬ Ｃｈｉｎａ。In addition, J, BioL China.
２５０４００７ （１９７５）にあるようにこの両者を
一体化するため、アガロースでかためるものもある。本
発明に係る２次元電気泳動法は基板（ガラス、ポリエス
テルなど）にシランカプリング剤を介してポリアクリル
アミドゲルの片面を化学的に結合させ、他の片面を開放
状態をしたいわゆるオープンゲルを用いた水平式泳動法
である。この場合２次元目の泳動に用いる濃度勾配ゲル
の低濃度側に設けた溝の中に短冊型の１次元電気泳動用
支持体を入れ両者をアガロースで一体化したのち通電す
る。2504007 (1975), in order to integrate the two, some are hardened with agarose. The two-dimensional electrophoresis method according to the present invention uses a so-called open gel in which one side of a polyacrylamide gel is chemically bonded to a substrate (glass, polyester, etc.) via a silane coupling agent, and the other side is left open. This is a horizontal electrophoresis method. In this case, a strip-shaped support for one-dimensional electrophoresis is placed in a groove provided on the low-concentration side of a concentration gradient gel used for second-dimensional electrophoresis, and after integrating the two with agarose, electricity is applied.
従来この操作は手作業で行なわれているが、これを自動
化する場合、このアガロースは前もって融解、保温して
液状に保ち、あらかじめ設定された時間に溝の中に注入
し、■、２次元泳動用支持体が滑らかに結合されるよう
にしなくては８ならない。Conventionally, this operation has been performed manually, but when automating this, the agarose is melted in advance, kept warm to keep it in a liquid state, and injected into the groove at a preset time. It must be ensured that the supports are joined together smoothly.
本発明の目的は自動２次元電気泳動装置において、１，
２次元泳動用支持体を一体化させるアガロースを必要と
される時間隔時状態で保ち、設定された位置に注入する
機構を備えた自動２次元電気泳動装置を提供することに
ある。An object of the present invention is to provide an automatic two-dimensional electrophoresis apparatus that includes:
An object of the present invention is to provide an automatic two-dimensional electrophoresis device having a mechanism for maintaining agarose integrated with a support for two-dimensional electrophoresis in a required time interval state and injecting it into a set position.
分離性能の優れた２次元電気泳動像を得るための要素の
一つに１次元泳動分離した試料の２次元支持体への移行
の条件がある。アガロースによる１次元泳動用ゲルと２
次元泳動用ゲルの結合はこれを左右する因子である。ア
ガロースによる結合に関して従来個々に手作業で°おこ
なっていた動作を自動化するには、アガロースの加熱保
温と所定の位置への注入動作の機構が必要になってくる
。One of the factors for obtaining a two-dimensional electrophoretic image with excellent separation performance is the conditions for transferring a sample separated by one-dimensional electrophoresis to a two-dimensional support. Gel for one-dimensional migration using agarose and 2
The binding of the dimensional gel is a factor that influences this. In order to automate the operations associated with binding with agarose, which were previously performed individually and manually, a mechanism for heating and keeping the agarose and injecting it into a predetermined position is required.
アガロースを融解させ、そのまま保持する温度は以下の
条件を満足させるように設定されなくてはならない６す
なわち、（１）沸とうさせない。（２）この装置は主と
して蛋白質試料の分析に用いられるものであるから支持
体中の蛋白質を変性させる程高温ではいけない。（３）
冷却板にのせである泳動用支持体は通常４℃近い低温に
しであるため、アガロースが１．２次元用ゲルの隙間に
充分浸入するまでは融けた状態を保つ。などである。他
方シリンジ付アガロース容器は移動台に取りつけると共
に直線移動用モータ、シリンジ駆動モータで所定の位置
へのアガロース注入をはかった。The temperature at which agarose is melted and maintained must be set so as to satisfy the following conditions6: (1) Do not allow it to boil. (2) Since this device is mainly used for analyzing protein samples, the temperature must not be high enough to denature the protein in the support. (3)
Since the electrophoresis support placed on the cooling plate is normally kept at a low temperature of around 4°C, it remains in a molten state until the agarose sufficiently penetrates into the gaps of the 1.2-dimensional gel. etc. On the other hand, the agarose container with a syringe was attached to a moving table, and agarose was injected into a predetermined position using a linear movement motor and a syringe drive motor.
以下本発明の一実施例を第１図および第２図により説明
する。An embodiment of the present invention will be described below with reference to FIGS. 1 and 2.
水平式オープンゲル２次元電気泳動法とは短冊型の基板
（ガラス、ポリエステルなど）に片面を結合させ、他の
片面を開放状態にしたポリアクリルアミドゲルを用いた
電気泳動法である。短冊型の１次元泳動用ゲル１を用い
て泳動分離したものを、同じく基板結合のポリアクリル
アミド濃度勾配ゲル２の一端に設けた溝の中に配置した
のち通電し、２次元展開させるものである。１次元泳動
分離した試料を２次元泳動用支持体の中に滑らかに移行
させるには、１，２次元ゲルの密着度が重要なポイント
となっている。 −一、
二次元ゲルを密着させるためには１次元用支持体を２次
元用支持体の溝に入れたのち、両者の隙間にアガロース
（たとえば低融点アガロースの１％溶液）を流し込んだ
のち固化して１体化するのが良い７本発明はこのプロセ
スの自動化に関するもので、あらかじめ調製、融解した
アガロースをシリンジ容器３に一定量吸引しておく、シ
リンジ針４を含む容器全体をシート状のヒータ６で包み
、絶縁トランス９．摺動抵抗器１０を通し、て適正温度
にコントロールする。適正温度とは沸騰せず、固化せず
、さらに上記２種類のゲルの隙間を滑らかに埋めたのち
出来るだけ早く固化する温度を云う。例えば、アガロー
スの製品によっても異なるが８０℃で用いる。尚シリン
ジの針先は最初２次元用ゲル中の溝に近い位置に置き、
アガロース液を少量ずつ押出しながら溝に沿って移動さ
せる必要がある。これはシリンジを移動台７に取りつけ
、軸１１をシリンジ駆動モータ１５で動かす動作とシリ
ンジ直線移動駆動モータ１８で移動させる構造で対処し
ている。なお熱効率をよくすると同時に周辺の温度を覚
乱させないためには。The horizontal open gel two-dimensional electrophoresis method is an electrophoresis method using a polyacrylamide gel with one side bonded to a strip-shaped substrate (glass, polyester, etc.) and the other side left open. The gel separated by electrophoresis using a strip-shaped one-dimensional electrophoresis gel 1 is placed in a groove provided at one end of a polyacrylamide concentration gradient gel 2, which is also bound to a substrate, and then energized to develop it two-dimensionally. . In order to smoothly transfer a sample separated by one-dimensional electrophoresis into a support for two-dimensional electrophoresis, the degree of adhesion between the one- and two-dimensional gels is an important point. -1,
In order to adhere the two-dimensional gel, the one-dimensional support is placed in the groove of the two-dimensional support, and agarose (for example, a 1% solution of low melting point agarose) is poured into the gap between the two and solidified. 7 The present invention relates to automation of this process, in which a fixed amount of pre-prepared and melted agarose is sucked into the syringe container 3, and the entire container including the syringe needle 4 is heated with a sheet-like heater 6. Wrapping, insulation transformer9. The temperature is controlled at an appropriate temperature through a sliding resistor 10. The appropriate temperature is the temperature at which the gel does not boil, does not solidify, and solidifies as quickly as possible after smoothly filling the gap between the two types of gels. For example, it is used at 80°C, although it varies depending on the agarose product. First, place the needle tip of the syringe near the groove in the two-dimensional gel.
It is necessary to move the agarose liquid along the groove while extruding it little by little. This is handled by a structure in which the syringe is mounted on a moving table 7, and the shaft 11 is moved by a syringe drive motor 15 and a syringe linear movement drive motor 18. In order to improve thermal efficiency and at the same time not disturb the surrounding temperature.
断熱材にて周囲を覆う必要がある。It is necessary to cover the surrounding area with insulation material.
シート状のヒータをアガロースを入れる容器に直接接触
させるため、効果よく温度コントロールすることが可能
となり、さらにコンパクトな装置とすることが出来る。Since the sheet-shaped heater is brought into direct contact with the container containing the agarose, it is possible to effectively control the temperature, and the apparatus can be made more compact.
第１図は水平式２次元電気泳動に用いる支持体の見取図
、第２図はアガロースの加熱機構を備えたアガロース注
入装置の見取図である。
ｌ・・弓次元泳動用の基板結合ポリアクリルアミドゲル
、２・・・２次元泳動用の基板結合ポリアクリルアミド
濃度勾配ゲル、３・・・シリンジ容器。
４・・・シリンジ針、５・・・シリンジ押棒、６・・・
ヒータ。
７・・・移動台、８・・・電源、９・・・＠縁トランス
。
ｌＯ・・・摺動抵抗器ｔ、 ｌ １・・・軸、１２・・
・ネジ。
１３・・・ナツト、１４・・・ガイド、１５・・・シリ
ンジ駆動モータ、１６・・・軸受、１７・・・軸、１８
・・・シリンジ直線移動駆動モータ。FIG. 1 is a sketch of a support used in horizontal two-dimensional electrophoresis, and FIG. 2 is a sketch of an agarose injection device equipped with an agarose heating mechanism. l... Substrate-bound polyacrylamide gel for bow-dimensional migration, 2... Substrate-bound polyacrylamide concentration gradient gel for two-dimensional migration, 3... Syringe container. 4...Syringe needle, 5...Syringe push rod, 6...
heater. 7...Moving table, 8...Power supply, 9...@rim transformer. lO...sliding resistor t, l1...shaft, 12...
·screw. 13... Nut, 14... Guide, 15... Syringe drive motor, 16... Bearing, 17... Shaft, 18
...Syringe linear movement drive motor.
Claims (1)
体と２次元泳動用支持体の結合材であるアガロースを融
解、保温するためアガロース用容器あるいはシリンジの
周囲にシート状のヒータを配置したことを特徴とする２
次元電気泳動装置。1. In an automatic two-dimensional electrophoresis device, a sheet-shaped heater is placed around the agarose container or syringe in order to melt and keep warm the agarose that is the bonding material between the one-dimensional electrophoresis support and the two-dimensional electrophoresis support. 2 characterized by
Dimensional electrophoresis device.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59225740A JPS61104247A (en) | 1984-10-29 | 1984-10-29 | 2D electrophoresis device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59225740A JPS61104247A (en) | 1984-10-29 | 1984-10-29 | 2D electrophoresis device |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS61104247A true JPS61104247A (en) | 1986-05-22 |
Family
ID=16834088
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP59225740A Pending JPS61104247A (en) | 1984-10-29 | 1984-10-29 | 2D electrophoresis device |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS61104247A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2011033547A (en) * | 2009-08-04 | 2011-02-17 | Hoyu Co Ltd | Two-dimensional electrophoretic method |
-
1984
- 1984-10-29 JP JP59225740A patent/JPS61104247A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2011033547A (en) * | 2009-08-04 | 2011-02-17 | Hoyu Co Ltd | Two-dimensional electrophoretic method |
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