Aquaporins extraction device

Abstract

The invention belongs to the technical field of aquaporin extraction, and particularly relates to an aquaporin extraction device which comprises a bottom driving plate, a rotating cup is mounted at the top of the bottom driving plate, a top cover is mounted at the top of the rotating cup, and at least one detachable connecting plate sheet is connected between the bottom driving plate and the top cover. A cup bin with an upward opening is arranged in the rotating cup, a supernatant collector is arranged at the position, close to the top end, in the cup bin, a bearing piece is fixed to the inner wall of the cup bin, a fixing piece is fixed to the position, located at the top of the bearing piece, of the outer side of the supernatant collector, and the fixing piece and the bearing piece are arranged in an attached mode. And a driving assembly is further installed in the bottom driving plate and used for driving the supernatant collector to rotate, and the driving assembly comprises an annular gear fixed to the inner wall of the top of the rotating cup. According to the invention, the supernatant can be conveniently collected after centrifugation, the pollution to the solution is avoided, the separation effect is ensured, and the equipment is simple to operate.

Description

Technical Field

[0001] The invention belongs to the technical field of water channel protein extraction, and in particular relates to a water channel protein extraction device. Background Art

[0002] In biochemistry and molecular biology research, the extraction and purification of aquaporins is an important step in studying the transmembrane transport mechanism of water molecules. Aquaporins are widely present in cell membranes and are responsible for regulating the water exchange inside and outside the cell. In order to study its structure and function, it is necessary to extract and obtain high-purity aquaporins from cells or tissues.

[0003] Existing methods for extracting aquaporins usually involve steps such as cell disruption and centrifugation. During the centrifugation process, cell fragments, unbroken cells, and target proteins are separated into different centrifuge cup layers. However, the traditional centrifuge cup design usually has only one cavity for holding liquid, which inevitably leads to some problems when collecting the supernatant after centrifugation.

[0004] First, after centrifugation is completed, when the centrifuge cup is opened or the supernatant is sucked out, the bacteria or cells that have not been completely centrifuged to the bottom may be resuspended due to vibration or disturbance, which will contaminate the supernatant and affect the subsequent separation effect. In addition, multiple transfers and transfers are required during the operation, which not only increases the complexity of the operation, but also may introduce more contamination risks, affecting the accuracy and repeatability of the experimental results.

[0005] Traditional centrifuge tubes require the use of a pipette to carefully aspirate the supernatant after centrifugation, but in this process, the sediment at the bottom is easily disturbed due to the inconvenience of operation. Although some centrifugal separation devices provide multi-layer designs, these layers are usually achieved through physical partitions, which limits the effective separation between layers.

[0006] Therefore, there is an urgent need for an improved aquaporin extraction device. Summary of the invention

[0007] The purpose of the present invention is to provide a water channel protein extraction device, which can conveniently collect the supernatant after centrifugation, avoid contamination of the solution, ensure the separation effect, and the device is simple to operate and ensures the accuracy and repeatability of the experimental results.

[0008] The technical solution adopted by the present invention is as follows:

[0009] A water channel protein extraction device comprises a bottom driving plate, a rotating cup is installed on the top of the bottom driving plate, a top cover is installed on the top of the rotating cup, at least one detachable connecting plate is connected between the bottom driving plate and the top cover, a cup bin with an upward opening is arranged inside the rotating cup, a supernatant collector is arranged inside the cup bin and near the top, a receiving sheet is fixed to the inner wall of the cup bin, a fixing sheet is fixed to the outer side of the supernatant collector and located on the top of the fixing sheet, and the fixing sheet is arranged in contact with the receiving sheet.

[0010] A driving assembly is also installed inside the bottom driving plate, and the driving assembly is used to drive the supernatant collector to rotate;

[0011] The driving assembly includes an annular gear fixed to the inner wall of the top of the rotating cup, and a driven gear is also arranged inside the rotating cup, and the driven gear is rotatably connected to the top cover, and the driven gear is meshingly connected to the annular gear, and a rotating gear is fixed to the outer side of the supernatant collector, and the rotating gear is meshingly connected to the driven gear.

[0012] The supernatant collector is provided with a supernatant collecting groove inside, a bottom groove with an opening facing downward is provided at the center position inside the supernatant collector, and the bottom groove is separated from the supernatant collecting groove by a spacer;

[0013] The inner walls of the supernatant collector and the spacer are both provided with first liquid leakage grooves at positions near the top, and the inner walls of the supernatant collector and the spacer are installed at positions corresponding to the first liquid leakage grooves with sealing mechanisms for sealing the first liquid leakage grooves.

[0014] The blocking mechanism includes a first mounting groove arranged in the inner wall of the supernatant collector and the spacer and located on one side of the first liquid leakage groove, a first blocking plate is slidably connected to the interior of the first mounting groove, and the first blocking plate slides toward one side of the first liquid leakage groove and extends into the first liquid leakage groove, a first spring is also installed inside the first mounting groove, and the first spring is connected to the first blocking plate.

[0015] A second liquid leakage groove is provided at the bottom of the supernatant collector, a second installation groove is provided on the inner wall of the bottom of the supernatant collector and on one side of the second liquid leakage groove, a second blocking plate is slidably connected inside the second installation groove, and the second blocking plate slides toward the side close to the second liquid leakage groove and extends into the second liquid leakage groove, a second spring is also installed inside the second installation groove, and the second spring is connected to the second blocking plate;

[0016] A recovery mechanism is also installed inside the second installation groove, and the recovery mechanism is used to recover the second blocking plate.

[0017] The recovery mechanism includes an electromagnet fixed inside the second mounting groove, and the electromagnet is installed on the inner wall of the second mounting groove away from the second blocking plate. A connecting magnet plate is fixed to the end of the second blocking plate close to the electromagnet, and the magnetic poles of the electromagnet and the connecting magnet plate on the side close to each other are opposite.

[0018] The technical effects achieved by the present invention are:

[0019] The invention can conveniently collect the supernatant after centrifugation, avoids contamination of the solution, ensures the separation effect, and the device is easy to operate, ensuring the accuracy and repeatability of the experimental results. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 It is a schematic diagram of the structure of the present invention;

[0021] Figure 2 It is a top view of the whole of the present invention;

[0022] Figure 3 It is a schematic diagram of the structure between the ring gear, the driven gear and the rotating gear in the present invention;

[0023] Figure 4 It is a schematic diagram of the structure between the driven gear and the magnet sheet in the present invention;

[0024] Figure 5 It is a schematic diagram of the structure between the fixing sheet and the receiving sheet in the present invention;

[0025] Figure 6 It is a schematic diagram of the structure between the supernatant collector and the first baffle in the present invention;

[0026] Figure 7 is a cross-sectional view of a supernatant collector in the present invention;

[0027] Figure 8 It is a schematic diagram of the structure between the supernatant collector, the first spring and the first blocking plate in the present invention;

[0028] Fig. 9 is a schematic diagram of the structure between the bottom groove and the second blocking plate in the present invention;

[0029] Fig.10 It is a schematic diagram of the structure between the second spring, the second blocking plate and the electromagnet in the present invention.

[0030] In the accompanying drawings, the components represented by the reference numerals are listed as follows:

[0031] 1. Bottom driving plate; 2. Rotating cup; 3. Cup chamber; 4. Ring gear; 5. Driven gear; 6. Rotating gear; 7. Supernatant collector; 8. Driving guide gear; 9. Magnet sheet; 10. Fixed sheet; 11. Receiver sheet; 12. Bottom groove; 13. Supernatant collecting groove; 14. First blocking plate; 15. First mounting groove; 16. First spring; 17. Second blocking plate; 18. Second mounting groove; 19. Second spring; 20. Connecting magnet plate; 21. Electromagnet; 22. Top cover; 23. Connecting plate; 24. Spacer; 25. First leakage groove; 26. Second leakage groove. DETAILED DESCRIPTION

[0032] In order to make the purpose and advantages of the present invention more clearly understood, the present invention is specifically described below in conjunction with embodiments. It should be understood that the following text is only used to describe one or several specific embodiments of the present invention, and does not strictly limit the scope of protection of the specific claims of the present invention.

[0033] like Figure 1-10 As shown, a water channel protein extraction device comprises a bottom driving plate 1, a rotating cup 2 is installed on the top of the bottom driving plate 1, a motor or other device capable of driving the rotating cup 2 to rotate is installed at the center position inside the bottom driving plate 1, when the motor is driven, it can drive the rotating cup 2 to rotate, a top cover 22 is installed on the top of the rotating cup 2, at least one detachable connecting plate 23 is connected between the bottom driving plate 1 and the top cover 22, and a cup bin 3 with an upward opening is arranged inside the rotating cup 2;

[0034] When extracting aquaporins, the following steps are taken:

[0035] Disrupting cells or tissues by physical methods such as ultrasonication, homogenization, or chemical methods such as enzymatic hydrolysis to release intracellular contents, including aquaporins;

[0036] Ultracentrifugation is used to separate cell membranes and other organelles. The precipitate after centrifugation contains cell membranes, which are an important source of aquaporins.

[0037] The pellet after centrifugation was dissolved in a buffer containing a nonionic surfactant to extract membrane proteins;

[0038] Purify the extracted membrane protein using affinity chromatography techniques, for example, using an affinity column that binds to a specific antibody to separate the target protein from the mixture;

[0039] Electrophoresis and detection: Use SDS-PAGE electrophoresis to separate the purified water channel protein, and detect it by Western Blot and other methods to verify the purity and content of the protein;

[0040] The above steps need to be performed at low temperatures to ensure protein stability and activity;

[0041] This device separates the cell membrane and other organelles by centrifugation, as follows:

[0042] Add the tissue sample into the cup chamber 3, start the preset motor inside the bottom driving plate 1, which can drive the rotating cup 2 to rotate, and drive the tissue sample inside the cup chamber 3 to rotate, so that the tissue sample can be centrifuged;

[0043] A supernatant collector 7 is arranged inside the cup bin 3 and near the top. A receiving sheet 11 is fixed to the inner wall of the cup bin 3. A fixing sheet 10 is fixed to the outside of the supernatant collector 7 and located on the top of the receiving sheet 11. The fixing sheet 10 is fitted with the receiving sheet 11. The receiving sheet 11 is used to receive the fixing sheet 10, so that the supernatant collector 7 can always be inside the cup bin 3 and will not fall off.

[0044] A driving assembly is also installed inside the bottom driving plate 1 , and the driving assembly is used to drive the clear liquid collector 7 to rotate.

[0045] Refer to the attached Figure 6-8 The supernatant collector 7 is provided with a supernatant collecting groove 13, and a bottom groove 12 with an opening facing downward is provided at the center of the supernatant collector 7, and the bottom groove 12 is separated from the supernatant collecting groove 13 by a spacer 24. The arrangement of the supernatant collecting groove 13 and the bottom groove 12 reduces the contact area between the supernatant collector 7 and the centrifuged solution, thereby not affecting the centrifugal effect;

[0046] The first liquid leakage groove 25 is provided on the inner wall of the supernatant collector 7 and the spacer 24 and near the top;

[0047] The supernatant collector 7 is placed inside the cup chamber 3, and the supernatant produced after centrifugation can be collected through the supernatant collector 7 and the first leakage groove 25 provided on the inner wall of the spacer 24. The supernatant part can be removed, and the remaining part including the precipitate part containing an important source of aquaporin is kept for standby;

[0048] A blocking mechanism is installed on the inner wall of the supernatant collector 7 and the spacer 24 at a position corresponding to the first liquid leakage groove 25 , and the blocking mechanism is used to block the first liquid leakage groove 25 .

[0049] See attached Figure 8As shown, the blocking mechanism includes a first mounting groove 15 arranged in the inner wall of the supernatant collector 7 and the spacer 24 and located on one side of the first liquid leakage groove 25, and a first blocking plate 14 is slidably connected inside the first mounting groove 15. After being subjected to the centrifugal rotation force, the first blocking plate 14 is subjected to the centrifugal force and overcomes the elastic force of the first spring 16, and gradually moves toward the inside of the first mounting groove 15, and the first blocking plate 14 slides toward one side of the first liquid leakage groove 25 and extends into the first liquid leakage groove 25, and a first spring 16 is also installed inside the first mounting groove 15, and the first spring 16 is connected to the first blocking plate 14;

[0050] Refer to the attached Figure 3 The driving assembly includes a ring gear 4 fixed to the inner wall of the top of the rotating cup 2, a driven gear 5 is also provided inside the rotating cup 2, and the driven gear 5 is rotatably connected to the top cover 22, and the driven gear 5 is meshed with the ring gear 4, and a rotating gear 6 is fixed to the outer side of the clear water collector 7, and the rotating gear 6 is meshed with the driven gear 5;

[0051] When the rotating cup 2 rotates, the ring gear 4 can be driven to rotate, and the ring gear 4 can drive the driven gear 5 to rotate through the meshing connection between the ring gear 4 and the driven gear 5, and the driven gear 5 is rotatably connected with the top cover 22, so that the driven gear 5 can rotate and drive the rotating gear 6 and the supernatant collector 7 to rotate, so that the supernatant collector 7 can rotate, and the rotation speed of the supernatant collector 7 is different from the rotation speed of the rotating cup 2;

[0052] See attached Figure 4 , a plurality of magnet pieces 9 are fixed inside the rotating cup 2 and at the bottom of the driven gear 5. Through the arrangement of the magnet pieces 9, when the driven gear 5 stops rotating, the tooth part of the driven gear 5 can stay and be adsorbed on the magnet pieces 9 according to the arrangement of the magnet pieces 9, so that when the supernatant collector 7 is taken out and placed inside the cup bin 3, the driven gear 5 can mesh with the teeth of the rotating gear 6. The material of the driven gear 5 can be iron. A plurality of driving guide teeth 8 are fixed on the inner wall of the cup bin 3 and away from the side of the driven gear 5, and the plurality of driving guide teeth 8 are located at the oblique upper part of the driven gear 5. Through this arrangement, the driving guide teeth 8 guide the rotating gear 6, so that the rotating gear 6 and the driven gear 5 can be just meshed after installation;

[0053] When the rotating cup 2 rotates, the supernatant collector 7 also rotates accordingly. The supernatant collector 7 drives the first blocking plate 14 back to the first mounting groove 15 through its own centrifugal force, so that the supernatant can enter the supernatant collecting groove 13 through the first leakage groove 25. The above-mentioned drive assembly is set so that the rotating cup 2 and the supernatant collector 7 have different rotation speeds, so that during centrifugation, the supernatant can better enter the supernatant collecting groove 13, and the effect of the supernatant entering the supernatant collecting groove 13 will not be limited due to the same speed.

[0054] The first blocking plate 14 can be made of metal such as steel, and can enter the first installation groove 15 through its own weight and the centrifugal effect. After the centrifugation is completed, the first blocking plate 14 can rely on the setting of the first spring 16, so that the first spring 16 bounces the first blocking plate 14 back into the first leakage groove 25 to block the first leakage groove 25.

[0055] See attached Figure 9-10 A second liquid leakage groove 26 is provided at the bottom of the supernatant collector 7, and a second mounting groove 18 is provided on the inner wall of the bottom of the supernatant collector 7 and on one side of the second liquid leakage groove 26. A second blocking plate 17 is slidably connected inside the second mounting groove 18, and the second blocking plate 17 slides toward the side close to the second liquid leakage groove 26 and extends into the second liquid leakage groove 26. The second blocking plate 17 can block the second liquid leakage groove 26. A second spring 19 is also installed inside the second mounting groove 18, and the second spring 19 is connected to the second blocking plate 17;

[0056] With this arrangement, the second spring 19 can be arranged so that the second spring 19 can always bounce the second blocking plate 17 to be located in the second liquid leakage groove 26, thereby blocking the second liquid leakage groove 26;

[0057] A recovery mechanism is also installed inside the second installation groove 18 , and the recovery mechanism is used to recover the second blocking plate 17 .

[0058] See attached Fig.10 The recovery mechanism includes an electromagnet 21 fixed inside the second mounting groove 18, and the electromagnet 21 is installed on the inner wall of the second mounting groove 18 away from the second blocking plate 17. A connecting magnet plate 20 is fixed to one end of the second blocking plate 17 close to the electromagnet 21, and the magnetic poles of the electromagnet 21 and the connecting magnet plate 20 close to each other are opposite;

[0059] When the supernatant is collected more and the lower sediment is collected together, the electromagnet 21 can be driven to emit magnetism, so that the electromagnet 21 can attract the connecting magnet plate 20, so that the second leakage groove 26 is no longer blocked by the second blocking plate 17, so that the sediment collected in the supernatant collection groove 13 flows into the cup chamber 3 through the second leakage groove 26;

[0060] The cup chamber 3 is provided with a scale inside, and the user can add a corresponding volume of liquid into the cup chamber 3 according to the scale, so that the boundary line between the supernatant part and the sediment part is roughly equivalent to the position of the first leakage groove 25 on the supernatant collector 7, thereby reducing the situation where the second leakage groove 26 discharges part of the solution;

[0061] Based on this, the application enables convenient collection of the supernatant after centrifugation.

[0062] The above is only a preferred embodiment of the present invention. It should be noted that, for those skilled in the art, several improvements and modifications can be made without departing from the principles of the present invention, and these improvements and modifications should also be considered as the protection scope of the present invention. The structures, devices and operating methods not specifically described and explained in the present invention shall be implemented according to the conventional means in the art unless otherwise specified and limited.

Claims

1. A water channel protein extraction device, comprising a bottom driving plate (1), characterized in that: A rotating cup (2) is installed on the top of the bottom driving plate (1), a top cover (22) is installed on the top of the rotating cup (2), at least one detachable connecting plate (23) is connected between the bottom driving plate (1) and the top cover (22), a cup bin (3) with an upward opening is arranged inside the rotating cup (2), a supernatant collector (7) is arranged inside the cup bin (3) and near the top, a receiving plate (11) is fixed to the inner wall of the cup bin (3), a fixing plate (10) is fixed to the outer side of the supernatant collector (7) and located on the top of the fixing plate (11), and the fixing plate (10) is arranged in close contact with the fixing plate (11).

2. A water channel protein extraction device according to claim 1, characterized in that: A driving assembly is also installed inside the bottom driving plate (1), and the driving assembly is used to drive the supernatant collector (7) to rotate; The driving assembly comprises a ring gear (4) fixed to the inner wall of the top of the rotating cup (2); a driven gear (5) is also arranged inside the rotating cup (2); the driven gear (5) is rotatably connected to the top cover (22); the driven gear (5) is meshingly connected to the ring gear (4); a rotating gear (6) is fixed to the outer side of the supernatant collector (7); the rotating gear (6) is meshingly connected to the driven gear (5).

3. The water channel protein extraction device according to claim 1, characterized in that: A supernatant collecting groove (13) is arranged inside the supernatant collector (7), a bottom groove (12) with an opening facing downward is arranged at the center position inside the supernatant collector (7), and the bottom groove (12) and the supernatant collecting groove (13) are separated by a spacer (24); A first liquid leakage groove (25) is provided on the inner wall of the supernatant collector (7) and the spacer (24) at a position close to the top, and a blocking mechanism is installed on the inner wall of the supernatant collector (7) and the spacer (24) at a position corresponding to the first liquid leakage groove (25), and the blocking mechanism is used to block the first liquid leakage groove (25).

4. A water channel protein extraction device according to claim 3, characterized in that: The blocking mechanism comprises a first mounting groove (15) arranged in the inner wall of the supernatant collector (7) and the spacer (24) and located on one side of the first liquid leakage groove (25); a first blocking plate (14) is slidably connected inside the first mounting groove (15), and the first blocking plate (14) slides toward one side of the first liquid leakage groove (25) and extends into the first liquid leakage groove (25); a first spring (16) is also installed inside the first mounting groove (15), and the first spring (16) is connected to the first blocking plate (14).

5. The water channel protein extraction device according to claim 1, characterized in that: A second liquid leakage groove (26) is arranged at the bottom of the supernatant collector (7); a second mounting groove (18) is arranged on the inner wall of the bottom of the supernatant collector (7) and located on one side of the second liquid leakage groove (26); a second blocking plate (17) is slidably connected inside the second mounting groove (18); the second blocking plate (17) slides toward the side close to the second liquid leakage groove (26) and extends into the second liquid leakage groove (26); a second spring (19) is also installed inside the second mounting groove (18), and the second spring (19) is connected to the second blocking plate (17); A recovery mechanism is also installed inside the second installation groove (18), and the recovery mechanism is used to recover the second blocking plate (17).

6. The water channel protein extraction device according to claim 5, characterized in that: The recovery mechanism comprises an electromagnet (21) fixed inside the second mounting groove (18), and the electromagnet (21) is mounted on the inner wall of the second mounting groove (18) on a side away from the second blocking plate (17), a connecting magnet plate (20) is fixed on one end of the second blocking plate (17) close to the electromagnet (21), and the magnetic poles of the electromagnet (21) and the connecting magnet plate (20) on the sides close to each other are opposite.

7. The water channel protein extraction device according to claim 2, characterized in that: A plurality of magnet pieces (9) are fixed inside the rotating cup (2) and at the bottom of the driven gear (5).

8. The water channel protein extraction device according to claim 2, characterized in that: A plurality of driving guide teeth (8) are fixed on the inner wall of the cup bin (3) and on a side away from the driven gear (5), and the plurality of driving guide teeth (8) are located at an oblique upper portion of the driven gear (5).

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