CA2837319A1 - Compartmentalized gel-electrophoresis apparatus - Google Patents
Compartmentalized gel-electrophoresis apparatus Download PDFInfo
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- CA2837319A1 CA2837319A1 CA2837319A CA2837319A CA2837319A1 CA 2837319 A1 CA2837319 A1 CA 2837319A1 CA 2837319 A CA2837319 A CA 2837319A CA 2837319 A CA2837319 A CA 2837319A CA 2837319 A1 CA2837319 A1 CA 2837319A1
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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
- G01N27/44773—Multi-stage electrophoresis, e.g. two-dimensional 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
- G01N27/44704—Details; Accessories
- G01N27/44713—Particularly adapted electric power supply
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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
- G01N27/44704—Details; Accessories
- G01N27/44708—Cooling
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Abstract
An apparatus for performing gel electrophoresis using a plurality of cassette sockets that can accommodate a plurality of gel cassettes. The apparatus allows each gel cassette to be run concurrently and individually. In accordance to the suggested enhancements, different gel compositions and different liquid buffer solutions could be used simultaneously. The apparatus also permits a group of users to use the same apparatus at the same time for distinctive experiments. The electric current for each cassette can be switched on or off at any time without disturbing the progress in other subunits. A heat exchanger within the apparatus comprises of tubes that carry the coolant into the apparatus through an inlet and out through an outlet.
Description
Compartmentalized gel-electrophoresis apparatus Field of the invention:
The present invention relates to electrophoresis and more particularly to a design that improves the versatility and convenience of using electrophoretic devices.
Background:
Electrophoresis is a commonly employed technique that applies an electric field to separate desired molecules based on some of their properties, which includes (but is not necessarily limited to) their size, charge or binding affinity. In particular, electrophoresis of multiple samples is normally performed on a porous gel medium. The gel medium is normally immersed in a liquid (and in particular, buffers for biological applications). During electrophoresis, a current is run through the apparatus and forces molecules to migrate from one side of the gel to another. Each type of molecule will migrate at a different rate based on their chemical properties. Gels are usually run with a reference ladder to identify the size of the loaded molecules after the electrophoretic process.
Normally, multiple gels are run with the same conditions within the same container. One cannot stop the gel when it has already started running if one wants to add more samples to the same gel.
Doing so would make the comparison of the newly loaded samples to the reference ladder inaccurate.
Another common phenomenon is that some experiments (e.g. protein purifications) take multiple days to complete and something could go wrong early on in the experiment before you can check.
If left unchecked until near the end of the experiment, this could end up wasting time, money and reagents.
Rather than waiting until the end of the experiment to perform electrophoresis, one can perform electrophoresis more in "real time". In other words, one can simply start the electrophoretic process after obtaining a few samples, rather than waiting until near the end of the experiment where one has a lot more samples. In addition, one can also run multiple gels independently and can start or stop the electrophoretic process at any time without interrupting the other gel runs within the main electrophoretic unit. Furthermore, one can run multiple gels with different compositions or different running buffers, thereby eliminating the need to use extra space or additional electrophoresis units.
Prior art:
The current improvements on gel electrophoresis apparatuses are intended to accommodate a different scope of experiments. Examples of such improvements are:
1) EP 0644420 A2, Method and apparatus for gel electrophoresis Inventors: Chand ran R. Sabanayagam, George M. Holzwarth, Eric H. Lai The invention employs two or more electric fields in a single gel. According to this invention, which is defined as Multiple-Zone Pulsed-Field Gel Electrophoresis (MZPFGE), the electric field is set to different values in two or more spatially distinct regions of the gel.
The present invention relates to electrophoresis and more particularly to a design that improves the versatility and convenience of using electrophoretic devices.
Background:
Electrophoresis is a commonly employed technique that applies an electric field to separate desired molecules based on some of their properties, which includes (but is not necessarily limited to) their size, charge or binding affinity. In particular, electrophoresis of multiple samples is normally performed on a porous gel medium. The gel medium is normally immersed in a liquid (and in particular, buffers for biological applications). During electrophoresis, a current is run through the apparatus and forces molecules to migrate from one side of the gel to another. Each type of molecule will migrate at a different rate based on their chemical properties. Gels are usually run with a reference ladder to identify the size of the loaded molecules after the electrophoretic process.
Normally, multiple gels are run with the same conditions within the same container. One cannot stop the gel when it has already started running if one wants to add more samples to the same gel.
Doing so would make the comparison of the newly loaded samples to the reference ladder inaccurate.
Another common phenomenon is that some experiments (e.g. protein purifications) take multiple days to complete and something could go wrong early on in the experiment before you can check.
If left unchecked until near the end of the experiment, this could end up wasting time, money and reagents.
Rather than waiting until the end of the experiment to perform electrophoresis, one can perform electrophoresis more in "real time". In other words, one can simply start the electrophoretic process after obtaining a few samples, rather than waiting until near the end of the experiment where one has a lot more samples. In addition, one can also run multiple gels independently and can start or stop the electrophoretic process at any time without interrupting the other gel runs within the main electrophoretic unit. Furthermore, one can run multiple gels with different compositions or different running buffers, thereby eliminating the need to use extra space or additional electrophoresis units.
Prior art:
The current improvements on gel electrophoresis apparatuses are intended to accommodate a different scope of experiments. Examples of such improvements are:
1) EP 0644420 A2, Method and apparatus for gel electrophoresis Inventors: Chand ran R. Sabanayagam, George M. Holzwarth, Eric H. Lai The invention employs two or more electric fields in a single gel. According to this invention, which is defined as Multiple-Zone Pulsed-Field Gel Electrophoresis (MZPFGE), the electric field is set to different values in two or more spatially distinct regions of the gel.
2) EP 1230258 Al, Multi-compartment electrophoresis Inventors: Ben Herbert, Pier Giorgio Righetti A multi-compartment electrolyser design is used to prefractionate complex protein mixtures, for use prior to the implementation of 2-D maps; it helps to remove proteins present in large excess in a cell lysate or in body fluids.
3) WO 1992016830 Al, Segmented electrophoretic separation system useful for lipid profiling Inventors: Technology Inc Assay, Joyce Chang, Charles R Manning, Leroy 1 Pinto The device comprises of a plurality of gel matrixes, defining a liquid pathway upon which a directional electrophoretic gradient can be imposed. The gel matrices are in serial relationship and preferably are of a successively decreasing pore size.
4) WO 2005098408 Al, Multi function gel electrophoresis and apparatus Inventors: Charles B Scott, John Victor Mcknight, Steven., Elliott The invention enables the user to perform electrophoretic separation and electroblotting in the same system. Also provided are means to efficiently exchange heat within the system.
5) US 20040195103 Al, Vertical slab gel electrophoresis cell and method therefor Inventor: Deming Zhou An improved vertical slab gel electrophoresis cell and method for performing electrophoresis vertically within uprightly oriented slab shaped gel matrixes.
While these references may be adequate for their intended purpose, there is still a need for an apparatus that allows compartmentalization of gel cassettes with different gel conditions and different buffers during electrophoresis. There is also a need to simultaneously and independently run different gel cassettes, and to allow the addition and removal of gel cassettes without disturbing the electrophoresis process of other gel cassettes. All the features detailed in the prior arts are compatible with the proposed invention, including the advantage of the plurality of slots (which is exclusive to our apparatus). Therefore, there is still a need for improving the electrophoretic apparatus design to make the electrophoretic device more versatile.
Summary:
The factors noted in the background suggest that the current design of electrophoretic apparatus is robust. However, the design lacks versatility to accommodate the problems mentioned in the background. The present invention allows separate subunits (also referred to as "cassettes" in the invention description), in the main electrophoretic apparatus to be run simultaneously and independently. The addition or removal of one or more subunits at any time will not disturb the progress of other subunits that are within the main apparatus. It is also not mandatory to completely occupy every subunit slot ("cassette socket") in the main apparatus, as the electrophoretic process will still run properly. These features of the present invention also allow multiple users to share the main electrophoretic apparatus without consuming additional space (e.g. extra power supply units and extra electrophoretic chambers) because each subunit is independently run.
Gels with different compositions could be run in each subunit with different liquid buffer solutions (depending on the person operating the gel and what type of experiment is performed). Since each subunit can contain a small volume of liquid buffer solution, less buffer will be used per run (as opposed to electrophoretic runs in the current commercial electrophoretic chambers). However, smaller volumes of buffer will usually be unable to mitigate excessive heating of the gel during electrophoresis, so a heat exchanger is usually necessary for cooling the subunits during electrophoresis. This heat exchanger will allow the flow of a desired fluid (e.g. water) through the back of the main electrophoretic apparatus.
Additional features and advantages of the present invention will become more apparent from the detailed description that follows, taken in conjunction with the accompanying drawings.
Brief Description of Drawings Figure 1 is a front view of the present invention's main apparatus.
Figure 2 is a side view of the present invention's main apparatus.
Figure 3 is a front view of the subunit that accompanies the main apparatus.
Figure 4 is a side view of the subunit that accompanies the main apparatus.
Figure 5 is a top view of the subunit that accompanies the main apparatus.
Description While the features and principles that characterize this invention and distinguish it over the prior art may be implemented in a variety of ways and embodied in a variety of constructions, the invention as a whole can best be understood by examination of a specific example. One such example is depicted in the drawings.
Figure 1 depicts the front view of the main electrophoretic apparatus 1 of the invention. This apparatus can accommodate a plurality of slots, cassette sockets 3, that can accommodate gel cassettes 14. These gel cassettes are further depicted in Figure 3, Figure 4 and Figure 5. The main electrophoretic apparatus 1 comprises of a lid 2, an anode 6 and a cathode 7, both of which are attached to the lid 2 and connected to a power supply 8. The cathode carries the negative charge and the anode carries the positive charge. While the power supply is active, electrons enter through the cathode 7, travel through the cathode conductive wire 4 and into any inserted gel cassettes via the current conduction strips 5. The electrons will then exit into the anode conductive wire 11 and exit through the anode 6. Because an active power supply will produce heat throughout the system, a coolant will flow into the coolant inlet 9 and out of the coolant outlet 10 during electrophoresis.
The heat exchanger design is illustrated further in a side view of the main electrophoretic apparatus in Figure 2. There are a series of tubes that carries the coolant into the apparatus: the coolant inlet 9 and tubes 12 that carry coolant into the plane of the page, and the coolant outlet 10 and tubes 13 that carry coolant in the opposite direction of the inlet tubes (i.e. out of the plane of the page). In other words, pipes that carry coolant into the page are marked by a bold "X" in Figure 2, while pipes that carry coolant out of the plane of the page are marked by a bold dot in Figure 2.
Figure 2 also illustrates that the cassette socket 3 does not extend all the way from the front to the back of the main electrophoretic apparatus 1. Instead, the cassette socket only occupies part of the effective side dimension of the main apparatus. The rest of the side dimension is occupied by the heat exchanger.
Figure 3 depicts the front view of the gel cassette 14 of the invention that is intended to fit into the cassette sockets 3 (mentioned in Figure 1). A gel 15 with three gel wells 16 is placed into the gel cassette. The gel is held together by a back glass gel plate 19 and a front glass gel plate 20 as shown in Figure 4. The gel plates are securely held in place with a holder 22.
The gels in each cassette could be of different composition.
The gel cassette has a conductive strip 18 attached to the bottom of the cassette and attached to the gel cassette lid 17. The conductive strip 18 connects to the current conductive strips 5 when the gel cassette is inserted into the cassette sockets of the main electrophoretic apparatus. The conductive strip 18 actually extends a bit deeper into the gel cassette as shown in Figure 4. This ensures that the electric current will flow through the entire gel cassette and gel, so long as there is enough buffer to make contact with both the top and bottom conductive strips. The inside of the gel cassette is then filled with liquid buffer solution 21. The liquid buffer solutions can differ in composition among each gel cassette. Figure 5 further is a top view of the gel cassette and further helps in conceptualizing the gel cassette design.
While these references may be adequate for their intended purpose, there is still a need for an apparatus that allows compartmentalization of gel cassettes with different gel conditions and different buffers during electrophoresis. There is also a need to simultaneously and independently run different gel cassettes, and to allow the addition and removal of gel cassettes without disturbing the electrophoresis process of other gel cassettes. All the features detailed in the prior arts are compatible with the proposed invention, including the advantage of the plurality of slots (which is exclusive to our apparatus). Therefore, there is still a need for improving the electrophoretic apparatus design to make the electrophoretic device more versatile.
Summary:
The factors noted in the background suggest that the current design of electrophoretic apparatus is robust. However, the design lacks versatility to accommodate the problems mentioned in the background. The present invention allows separate subunits (also referred to as "cassettes" in the invention description), in the main electrophoretic apparatus to be run simultaneously and independently. The addition or removal of one or more subunits at any time will not disturb the progress of other subunits that are within the main apparatus. It is also not mandatory to completely occupy every subunit slot ("cassette socket") in the main apparatus, as the electrophoretic process will still run properly. These features of the present invention also allow multiple users to share the main electrophoretic apparatus without consuming additional space (e.g. extra power supply units and extra electrophoretic chambers) because each subunit is independently run.
Gels with different compositions could be run in each subunit with different liquid buffer solutions (depending on the person operating the gel and what type of experiment is performed). Since each subunit can contain a small volume of liquid buffer solution, less buffer will be used per run (as opposed to electrophoretic runs in the current commercial electrophoretic chambers). However, smaller volumes of buffer will usually be unable to mitigate excessive heating of the gel during electrophoresis, so a heat exchanger is usually necessary for cooling the subunits during electrophoresis. This heat exchanger will allow the flow of a desired fluid (e.g. water) through the back of the main electrophoretic apparatus.
Additional features and advantages of the present invention will become more apparent from the detailed description that follows, taken in conjunction with the accompanying drawings.
Brief Description of Drawings Figure 1 is a front view of the present invention's main apparatus.
Figure 2 is a side view of the present invention's main apparatus.
Figure 3 is a front view of the subunit that accompanies the main apparatus.
Figure 4 is a side view of the subunit that accompanies the main apparatus.
Figure 5 is a top view of the subunit that accompanies the main apparatus.
Description While the features and principles that characterize this invention and distinguish it over the prior art may be implemented in a variety of ways and embodied in a variety of constructions, the invention as a whole can best be understood by examination of a specific example. One such example is depicted in the drawings.
Figure 1 depicts the front view of the main electrophoretic apparatus 1 of the invention. This apparatus can accommodate a plurality of slots, cassette sockets 3, that can accommodate gel cassettes 14. These gel cassettes are further depicted in Figure 3, Figure 4 and Figure 5. The main electrophoretic apparatus 1 comprises of a lid 2, an anode 6 and a cathode 7, both of which are attached to the lid 2 and connected to a power supply 8. The cathode carries the negative charge and the anode carries the positive charge. While the power supply is active, electrons enter through the cathode 7, travel through the cathode conductive wire 4 and into any inserted gel cassettes via the current conduction strips 5. The electrons will then exit into the anode conductive wire 11 and exit through the anode 6. Because an active power supply will produce heat throughout the system, a coolant will flow into the coolant inlet 9 and out of the coolant outlet 10 during electrophoresis.
The heat exchanger design is illustrated further in a side view of the main electrophoretic apparatus in Figure 2. There are a series of tubes that carries the coolant into the apparatus: the coolant inlet 9 and tubes 12 that carry coolant into the plane of the page, and the coolant outlet 10 and tubes 13 that carry coolant in the opposite direction of the inlet tubes (i.e. out of the plane of the page). In other words, pipes that carry coolant into the page are marked by a bold "X" in Figure 2, while pipes that carry coolant out of the plane of the page are marked by a bold dot in Figure 2.
Figure 2 also illustrates that the cassette socket 3 does not extend all the way from the front to the back of the main electrophoretic apparatus 1. Instead, the cassette socket only occupies part of the effective side dimension of the main apparatus. The rest of the side dimension is occupied by the heat exchanger.
Figure 3 depicts the front view of the gel cassette 14 of the invention that is intended to fit into the cassette sockets 3 (mentioned in Figure 1). A gel 15 with three gel wells 16 is placed into the gel cassette. The gel is held together by a back glass gel plate 19 and a front glass gel plate 20 as shown in Figure 4. The gel plates are securely held in place with a holder 22.
The gels in each cassette could be of different composition.
The gel cassette has a conductive strip 18 attached to the bottom of the cassette and attached to the gel cassette lid 17. The conductive strip 18 connects to the current conductive strips 5 when the gel cassette is inserted into the cassette sockets of the main electrophoretic apparatus. The conductive strip 18 actually extends a bit deeper into the gel cassette as shown in Figure 4. This ensures that the electric current will flow through the entire gel cassette and gel, so long as there is enough buffer to make contact with both the top and bottom conductive strips. The inside of the gel cassette is then filled with liquid buffer solution 21. The liquid buffer solutions can differ in composition among each gel cassette. Figure 5 further is a top view of the gel cassette and further helps in conceptualizing the gel cassette design.
6 The invention can be used for multiple applications. One mode of carrying out the invention would be with SDS-PAGE verification of a protein purification process. Aliquots at each stage of the protein purification process will be taken and ran via SDS-PAGE to confirm that the protein purification was successful. Protein purification processes often require more than one day to complete. There is always a possibility of human error occurring during the protein purification process. If there is an error early in the process and this error is not caught early on, one can waste reagents and time.
Protein purification processes often use affinity chromatography in conjunction with a fusion protein. The fusion protein typically contains the protein of interest attached to a protein tag ("fusion tag") that has high affinity for a certain type of chromatography column. Usually one would elute the protein after binding the protein to the column with an elution buffer. This elution buffer would contain a compound that has a higher affinity for the chromatography column than the protein. One example of an error that could occur is if the lab personnel used an incorrect elution buffer to elute the protein. Typically, one would cleave the fusion tag off after elution and this process takes place for 12 or more hours to ensure complete cleavage. The protein usually undergoes further purification, e.g. via fast protein liquid chromatography (FPLC) and FPLC runs consume a lot of buffer solution and time. If this error were caught on the first day instead of the second day, then the amount of wasted time and reagents would be substantially less.
Protein purification processes often use affinity chromatography in conjunction with a fusion protein. The fusion protein typically contains the protein of interest attached to a protein tag ("fusion tag") that has high affinity for a certain type of chromatography column. Usually one would elute the protein after binding the protein to the column with an elution buffer. This elution buffer would contain a compound that has a higher affinity for the chromatography column than the protein. One example of an error that could occur is if the lab personnel used an incorrect elution buffer to elute the protein. Typically, one would cleave the fusion tag off after elution and this process takes place for 12 or more hours to ensure complete cleavage. The protein usually undergoes further purification, e.g. via fast protein liquid chromatography (FPLC) and FPLC runs consume a lot of buffer solution and time. If this error were caught on the first day instead of the second day, then the amount of wasted time and reagents would be substantially less.
7
Claims (11)
1. A method for gel electrophoresis, comprising:
- a main electrophoretic apparatus comprised of a plurality of slots to accommodate smaller subunits; and - said subunits that accommodate a plurality of gels and liquid buffer solution;
- a main electrophoretic unit which applies a voltage difference across any number of inserted subunits; and - a design such that the electric current travelling through any of the subunits can be started or stopped at any time without disturbing other subunits within the main apparatus;
- a heat exchanger embedded within the main embodiment that allows heat transfer between the subunits and the fluid that flows through the heat exchanger
- a main electrophoretic apparatus comprised of a plurality of slots to accommodate smaller subunits; and - said subunits that accommodate a plurality of gels and liquid buffer solution;
- a main electrophoretic unit which applies a voltage difference across any number of inserted subunits; and - a design such that the electric current travelling through any of the subunits can be started or stopped at any time without disturbing other subunits within the main apparatus;
- a heat exchanger embedded within the main embodiment that allows heat transfer between the subunits and the fluid that flows through the heat exchanger
2. Use according to claim 1, wherein the gels within each subunit could be of different composition.
3. Use according to claim 1, wherein the buffer within each subunit could potentially be of different composition.
4. Use according to claim 1, wherein the main electrophoretic apparatus comprises of a cathode and an anode.
5. Use according to claim 1, wherein the voltage difference can be identical for all inserted subunits.
6. Use according to claim 1, wherein the walls that separate each subunit from one another are impermeable to gas and liquid.
7. Use according to claim 1, wherein the heat exchanger is not compulsory for use if the heat production over time is insufficiently high to damage the apparatus, subunits or contents within the subunits.
8. Use according to claim 1, wherein the heat exchanger is of a shell-and-tube design.
9. Use according to claim 1, wherein electrically conductive material lines the inner walls of the subunit to ensure proper flow of electricity from the cathode to the anode (when connected to the main apparatus).
10. Use according to claim 1, wherein electrically conductive material lines the base and the top cover of the subunit to ensure proper flow of electricity from the cathode to the anode (when connected to the main apparatus).
11. Use according to claim 1, wherein the walls that separate each subunit from one another are composed of material that does not conduct electricity.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA2837319A CA2837319A1 (en) | 2013-12-19 | 2013-12-19 | Compartmentalized gel-electrophoresis apparatus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA2837319A CA2837319A1 (en) | 2013-12-19 | 2013-12-19 | Compartmentalized gel-electrophoresis apparatus |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2837319A1 true CA2837319A1 (en) | 2015-06-19 |
Family
ID=53477169
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2837319A Abandoned CA2837319A1 (en) | 2013-12-19 | 2013-12-19 | Compartmentalized gel-electrophoresis apparatus |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA2837319A1 (en) |
-
2013
- 2013-12-19 CA CA2837319A patent/CA2837319A1/en not_active Abandoned
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FZDE | Dead |
Effective date: 20161221 |