CN107014885A - A kind of prefabricated film of the isoelectric focusing electrophoresis for being exclusively used in sample preparation - Google Patents
A kind of prefabricated film of the isoelectric focusing electrophoresis for being exclusively used in sample preparation Download PDFInfo
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- CN107014885A CN107014885A CN201710341372.4A CN201710341372A CN107014885A CN 107014885 A CN107014885 A CN 107014885A CN 201710341372 A CN201710341372 A CN 201710341372A CN 107014885 A CN107014885 A CN 107014885A
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- isoelectric focusing
- gradient gel
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- focusing electrophoresis
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- 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
- G01N27/44756—Apparatus specially adapted therefor
- G01N27/44795—Isoelectric focusing
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Abstract
A kind of prefabricated film of the isoelectric focusing electrophoresis for being exclusively used in sample preparation, including foreboard, pH gradient gel, comb, sealing mucous membrane, back plate, vertical edge strip, wherein pH gradient gel is immobilization pH gradient gel or on-fixed pH gradient gel, immobilization pH gradient gel is synthesized in the glass sandwich that front and rear panel is surrounded, pH gradient gel is combined with glass plate, the first dimension IEF in 2DE and the second dimension PAGE features are organically combined, meet the demand of high flux isoelectric focusing experiment, simplify IEF operating process, improve the concentration effect of low-abundance protein.
Description
Technical field
It is pre- particularly for the isoelectric focusing electrophoresis of sample preparation the present invention relates to biotechnology separation detection apparatus field
Glue piece.
Background technology
Two dimensional gel electrophore- sis (Two-dimensional electrophoresis, 2DE) technology is used as protein research
Important means, its critical role is still unshakable in a short time.O ' Farrell and Klose was independently proposed in 1975
Come, and turn into modern times 2DE basis, its principle is：First dimension carries out isoelectric focusing, according to isoelectric point protein isolate matter, second
Dimension carries out polyacrylamide gel electrophoresis (PAGE) in the presence of lauryl sodium sulfate (SDS) and reducing agent, according to albumen
Matter bulk of molecule (being proportional to molecular weight) is separated.
As the key link in 2DE technologies, isoelectric focusing is the new technology that middle nineteen sixties occur.Electricity is waited in recent years
Focusing technology has new progress, has developed rapidly as a ripe modern age biochemical test technology.It is special that the technology is utilized
Ampholytes buffer solution in the gel (conventional polyacrylamide gel) one pH gradient of manufacture, every kind of protein during electrophoresis
To migrate to (now the protein net charge be 0) at its isoelectric point (pI) identical pH, form a very narrow zone.
In early days, people are led to after direct current using adding carrier ampholyte in electrolytic cell, carrier just according to respective isoelectric point not
With and move, and gradually form one by anode to negative electrode pH value gradually increased pH gradient.But prolonging with focal time
Long, pH gradient is unstable, is also easy to produce cathodic drift.In addition, IEF (isoelectric focusing) experimental implementation of early stage is very cumbersome, reappearance
Difference.Due to itself progress for many years technically, wherein mainly immobilization pH gradient (IPG, immobilized pH
Gradient invention and improvement), IEF technologies are used widely.IPG invention substantially increases IEF reappearance, and energy
Enough increase applied sample amount, so as to improve sensitivity.
There is also following defect by 2DE using IEF as core：Limited dynamic range；Separate size difference in sample larger
Albumen, the energy of low-abundance protein (such as signal protein and transcription factor) and hydrophobin (embrane-associated protein and transmembrane protein)
Power is limited；Immobilized ph gradient strip includes the configuration of rehydration buffer using the aquation of progress 12 hours or so is preceding needed on market today
Etc. step, operating process is wasted time and energy, and hydration process is insufficient in the presence of being swelled, and adhesive tape is damaged, and bubble equivalent risk occurs, seriously
Influence isoelectric focusing result；When sample size is more, low applied sample amount has a strong impact on the progress of subsequent experimental, multiple batches of sample
Processing increases the otherness between batch, the reduction of reliable experiment result degree.
In view of the above-mentioned problems, the present invention proposes a kind of prefabricated film of isoelectric focusing electrophoresis for being exclusively used in sample preparation, the dress
Put simple in construction, practical, easy to operate, substantially increase quantity of sample handling, reduce operation difficulty.
The content of the invention
A kind of prefabricated film of the isoelectric focusing electrophoresis for being exclusively used in sample preparation, such as Fig. 1, prefabricated film includes：Foreboard 1,
PH gradient gel 2, comb 3, sealing mucous membrane 4, back plate 5, vertical edge strip 6, foreboard 1 is placed in back plate 5, and with sealing mucous membrane 4
One end of foreboard 1, back plate 5 is sealed, pH gradient gel 2 is enclosed positioned at the lower surface of foreboard 1 by glass material, the upper surface of back plate 5
Into glass plies gap in, comb 3 be placed in the corresponding other end of sealing mucous membrane 4, wherein pH gradient gel 2 is gel film
And the prefabricated film is globality structure.
First, the volume of gel film is much larger than adhesive tape, and several samples, the electrophoresis experiment of high applied sample amount can be carried out simultaneously；Its
Secondary, the prefabricated film of the globality is combined pH gradient gel with glass plate, innovatively by the first dimension IEF in 2DE and the second dimension
PAGE features organically blend, and realize distillations of the 2DE in experimental technique under the new situation, the wherein prefabricated film chi of isoelectric focusing electrophoresis
It is very little to be matched with small-sized, medium-sized and large-scale Vertial electrophorestic tank, provide another technical tool for vast laboratory staff；In addition, on
Stating the prefabricated film of preparative isoelectric focusing electrophoresis can use in conventional Vertial electrophorestic tank, very simple be operated, without picture
Like product needs the specific expensive instrument of special purchase, hydration process of the prefabricated film without immobilized ph gradient strip, it is to avoid complicated behaviour
Make process, directly can be used to the prefabricated film of globality structure test.
It is preferred that, the comb 3 is combed for tack.When only a kind of sample needs to carry out isoelectric focusing, combed using tack, can
Applied sample amount is greatly increased, the electrophoresis experiment batch of gross sample is reduced, shortens the electrophoresis cycle；
It is preferred that, the comb 3 is combed for one to multiple hole.When sample type is more, using porous comb, and can be according to sample kind
The quantity of class selects the comb of corresponding hole count, synchronizes the isoelectric focusing of Multi-example, reduces sample treatment batch, accelerates experiment
Process, also effectively reduce in multiple batches of processing sample causes the difference of each batch experimental result because electrophoresis environment changes,
Experimental result can contrast improved.
It is preferred that, the vertical edge strip is the vertical edge strip of glass or the vertical edge strip of plastics.
It is preferred that, the vertical edge strip thickness is 0.1mm~2mm.The thickness of vertical edge strip is bigger, corresponding glass plies
Between gap thickness it is bigger, the thickness of open ended pH gradient gel 2 becomes big, can be according to the thickness to different pH gradient gels 2
Demand flexibly changes the thickness of vertical edge strip.
It is preferred that, the preceding plate thickness is 0.1mm~6mm.
It is preferred that, the rear plate thickness is 0.1mm~6mm.
It is preferred that, pH gradient gel 2 is immobilization pH ladder in a kind of described prefabricated film of preparative isoelectric focusing electrophoresis
Spend gel or on-fixed pH gradient gel.The cost of on-fixed pH gradient gel is relatively low；Immobilization pH gradient gel electrophoresis
Electric fields uniform, performance is stable, and high resolution, pH gradient flexible adjustment, the concentration effect of low-abundance protein is improved.
It is preferred that, the prefabricated film of isoelectric focusing electrophoresis can be used for isoelectric focusing experiment or dielectrophoresis to test.
In the prefabricated film of isoelectric focusing electrophoresis, pH gradient gel 2 is made in glass plies.By what is configured
Solution is injected into glass plies, makes the coagulation forming in glass plies of pH gradient gel 2.Because the rigidity of glass is big, heat conduction
Property it is small, so deflection is small, the thickness of pH gradient gel 2 after shaping is uniform, and gel electrophoresis electric field is consistent, and performance is stable, electrophoresis
Experimental result reliability is high.
Brief description of the drawings：
Embodiment is further described below in conjunction with the accompanying drawings, wherein：
Fig. 1 is the assembling schematic diagram of the prefabricated film of isoelectric focusing electrophoresis of the present invention.
Fig. 2 is the structural blast schematic diagram of the prefabricated film of isoelectric focusing electrophoresis of the present invention.
Fig. 3 is the structural blast schematic diagram of isoelectric focusing electrophoresis pre-prepared colloid piece in the specific embodiment of the invention 2.
Primary structure number explanation
| 1 | 2 | 3 | 4 | 5 | 6 |
| Foreboard | PH gradient gel | Comb | Seal mucous membrane | Back plate | Vertical edge strip |
Following embodiment will further illustrate the present invention with reference to above-mentioned accompanying drawing.
Case 1 is embodied：
The prefabricated film of preparative isoelectric focusing electrophoresis, including foreboard 1, pH gradient gel 2, comb 3, sealing mucous membrane 4, back plate
5 and vertical edge strip 6.A kind of described prefabricated film size of preparative isoelectric focusing electrophoresis with it is small-sized, medium-sized and large-scale vertical
Electrophoresis tank is matched；Described pH gradient gel is made in glass plies；Described comb is ten holes comb；Described foreboard 1, after
Plate 5 is glass material, and thickness is respectively 1mm, 1mm；Vertical edge strip 6 is the vertical edge strip of glass, and thickness is 1mm；Described pH
Gradient gel 2 is immobilization pH gradient gel, the gel pH stable of immobilization, high resolution；Present embodiment is by pH ladders
Degree gel is combined with glass plate, that is to say and innovatively the first dimension IEF in 2DE and the second dimension PAGE features organically blend, realize
Distillations of the 2DE in experimental technique under the new situation, for vast Isoelecric focusing gel electrophoresis laboratory staff provides another technology
Instrument.In addition, the prefabricated film of preparative isoelectric focusing electrophoresis made is used in existing Vertial electrophorestic tank, operation is very
Simply, without needing specially to buy specific expensive instrument application as like product, quantity of sample handling be normal experiment 30~
80 times, This reduces both experimental cost, and greatly facilitate user.
Case 2 is embodied：
As shown in figure 3, the prefabricated film of preparative isoelectric focusing electrophoresis, including foreboard 1, pH gradient gel 2, comb 3, sealing
Mucous membrane 4, back plate 5 and vertical edge strip 6.A kind of described prefabricated film size of preparative isoelectric focusing electrophoresis with it is small-sized, medium-sized
And large-scale Vertial electrophorestic tank matching；Described pH gradient gel is made in glass plies；Described comb 3 is tack comb；Institute
The foreboard 1 stated, back plate 5 are glass material, and thickness is respectively 2mm, 2mm；Vertical edge strip 6 is the vertical edge strip of plastics, and thickness is
1.5mm；Described pH gradient gel 2 is non-immobilization pH gradient gel, is prepared simply, with low cost；Present embodiment be by
PH gradient gel is combined with glass plate, that is to say and innovatively the first dimension IEF in 2DE and the second dimension PAGE features organically blend,
Distillations of the 2DE in experimental technique under the new situation is realized, another technical tool is provided for vast dielectrophoresis laboratory staff.
In addition, the prefabricated film of preparative isoelectric focusing electrophoresis made is used in existing Vertial electrophorestic tank, very simple is operated,
Without needing specially to buy specific expensive instrument application as like product, experimental cost is reduced, and greatly facilitates user,
The operating procedure of experiment is less.
Case 3 is embodied：
The prefabricated film of preparative isoelectric focusing electrophoresis, including foreboard 1, pH gradient gel 2, comb 3, sealing mucous membrane 4, back plate
5 and vertical edge strip 6.A kind of described prefabricated film size of preparative isoelectric focusing electrophoresis with it is small-sized, medium-sized and large-scale vertical
Electrophoresis tank is matched；Described pH gradient gel is made in glass plies；Described comb is 12 holes comb；Described foreboard 1,
Back plate 5 is glass material, and thickness is respectively 1mm, 1mm；Vertical edge strip 6 is the vertical edge strip of glass, and thickness is 2mm；Described
PH gradient gel 2 is immobilization pH gradient gel, the gel pH stable of immobilization, high resolution；Present embodiment is by pH
Gradient gel is combined with glass plate, that is to say and innovatively the first dimension IEF in 2DE and the second dimension PAGE features organically blend, real
Distillations of the 2DE in experimental technique is under the new situation showed, another technical tool is provided for vast dielectrophoresis laboratory staff.Separately
Outside, the prefabricated film of preparative isoelectric focusing electrophoresis of making is used in existing Vertial electrophorestic tank, operates very simple, and
It is not required to as like product needs specially to buy specific expensive instrument application, quantity of sample handling is 40~95 times of normal experiment, this
Both experimental cost is reduced, and greatly facilitates user.
Embodiment described above only expresses the several embodiments of the present invention, and it describes more specific and detailed, but simultaneously
Therefore the limitation to the scope of the claims of the present invention can not be interpreted as.It should be pointed out that for one of ordinary skill in the art
For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the guarantor of the present invention
Protect scope.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (10)
1. a kind of prefabricated film of the isoelectric focusing electrophoresis for being exclusively used in sample preparation, including foreboard, pH gradient gel, comb, sealing
Mucous membrane, back plate, vertical edge strip, foreboard are placed in back plate, and are sealed one end of foreboard, back plate with sealing mucous membrane, pH gradient
Gel is located in the glass plies gap surrounded by the foreboard of glass material, back plate, and comb is placed in corresponding another of sealing mucous membrane
One end, it is characterised in that：PH gradient gel is that the gel film and prefabricated film being fixed in glass plies are globality structure.
2. the prefabricated film of isoelectric focusing electrophoresis as claimed in claim 1, it is characterised in that：Comb is tack comb.
3. the prefabricated film of isoelectric focusing electrophoresis as claimed in claim 1, it is characterised in that：Comb is one to multiple hole comb.
4. the prefabricated film of isoelectric focusing electrophoresis as claimed in claim 1, it is characterised in that：Vertical edge strip be the vertical edge strip of glass or
The vertical edge strip of plastics.
5. the prefabricated film of isoelectric focusing electrophoresis as claimed in claim 1, it is characterised in that：Vertical edge strip thickness be 0.1mm~
2mm。
6. the prefabricated film of isoelectric focusing electrophoresis as claimed in claim 1, it is characterised in that：Preceding plate thickness is 0.1mm~6mm.
7. the prefabricated film of isoelectric focusing electrophoresis as claimed in claim 1, it is characterised in that：Plate thickness is 0.1mm~6mm afterwards.
8. the prefabricated film of isoelectric focusing electrophoresis as claimed in claim 1, it is characterised in that：PH gradient gel is immobilization pH gradient
Gel or on-fixed pH gradient gel.
9. the prefabricated film of the isoelectric focusing electrophoresis as described in any in claim 1~8, it is characterised in that：Available for isoelectric focusing
Experiment or dielectrophoresis experiment.
10. the prefabricated film of isoelectric focusing electrophoresis as claimed in claim 1, it is characterised in that：PH gradient gel is surrounded in forward and backward plate
Glass sheet separation in coagulation forming.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710341372.4A CN107014885A (en) | 2017-05-16 | 2017-05-16 | A kind of prefabricated film of the isoelectric focusing electrophoresis for being exclusively used in sample preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710341372.4A CN107014885A (en) | 2017-05-16 | 2017-05-16 | A kind of prefabricated film of the isoelectric focusing electrophoresis for being exclusively used in sample preparation |
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| CN107014885A true CN107014885A (en) | 2017-08-04 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201710341372.4A Pending CN107014885A (en) | 2017-05-16 | 2017-05-16 | A kind of prefabricated film of the isoelectric focusing electrophoresis for being exclusively used in sample preparation |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108359177A (en) * | 2018-02-25 | 2018-08-03 | 柴建锋 | A kind of pre-prepared colloid polypropylene material and preparation method thereof |
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| WO1994014526A1 (en) * | 1992-12-22 | 1994-07-07 | Bertin & Cie | Multiple electrophoresis device and method for the migration and transfer of macromolecules |
| US5637202A (en) * | 1993-10-27 | 1997-06-10 | California Institute Of Technology | Porous electrophoresis sponges |
| US20040045829A1 (en) * | 2002-04-12 | 2004-03-11 | Nikolaus Ingenhoven | Cassette, system, and 2-D gel electrophoresis method for separating molecules |
| CN1790003A (en) * | 2005-12-15 | 2006-06-21 | 西安工业学院 | Apparatus and method for making gradient gel film for full-automatic electrophoresis |
| US20140332390A1 (en) * | 2013-05-11 | 2014-11-13 | Biozax, Inc. | Prefabricated, self-contained gel electrophoresis module |
| CN105021685A (en) * | 2014-04-17 | 2015-11-04 | 上海伯楷安生物科技有限公司 | Anti-seepage gel-electrophoresis precast gel shell |
| CN205517665U (en) * | 2016-04-19 | 2016-08-31 | 天津纽赛生物技术有限公司 | Exempt from to join in marriage prefabricated glue |
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2017
- 2017-05-16 CN CN201710341372.4A patent/CN107014885A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1994014526A1 (en) * | 1992-12-22 | 1994-07-07 | Bertin & Cie | Multiple electrophoresis device and method for the migration and transfer of macromolecules |
| US5637202A (en) * | 1993-10-27 | 1997-06-10 | California Institute Of Technology | Porous electrophoresis sponges |
| US20040045829A1 (en) * | 2002-04-12 | 2004-03-11 | Nikolaus Ingenhoven | Cassette, system, and 2-D gel electrophoresis method for separating molecules |
| CN1790003A (en) * | 2005-12-15 | 2006-06-21 | 西安工业学院 | Apparatus and method for making gradient gel film for full-automatic electrophoresis |
| US20140332390A1 (en) * | 2013-05-11 | 2014-11-13 | Biozax, Inc. | Prefabricated, self-contained gel electrophoresis module |
| CN105021685A (en) * | 2014-04-17 | 2015-11-04 | 上海伯楷安生物科技有限公司 | Anti-seepage gel-electrophoresis precast gel shell |
| CN205517665U (en) * | 2016-04-19 | 2016-08-31 | 天津纽赛生物技术有限公司 | Exempt from to join in marriage prefabricated glue |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108359177A (en) * | 2018-02-25 | 2018-08-03 | 柴建锋 | A kind of pre-prepared colloid polypropylene material and preparation method thereof |
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Application publication date: 20170804 |
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