CN209858588U - Antibody incubation box for automatic western blotting method - Google Patents
Antibody incubation box for automatic western blotting method Download PDFInfo
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- CN209858588U CN209858588U CN201821843301.0U CN201821843301U CN209858588U CN 209858588 U CN209858588 U CN 209858588U CN 201821843301 U CN201821843301 U CN 201821843301U CN 209858588 U CN209858588 U CN 209858588U
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Abstract
The utility model relates to an automatic western blotting antibody incubation box, which comprises a box cover (1) and a box body (2), wherein a liquid storage device is arranged above the box cover (1), a recovery liquid storage device is arranged below the box body (2) and is arranged in a shell (3) with refrigeration and light-shielding functions, a plurality of clapboards (4) are arranged in the box body (2) to divide the box body into a plurality of PVDF film storage cavities (7); the liquid accumulator above the box cover (1) is connected with the top of the box cover (1), and the liquid accumulator below the box body (2) is connected with the bottom of the box body (2); the box cover (1) is provided with an anti-adding hole, a TBTS adding hole and an anti-adding hole, and the bottom of the box body (2) is provided with an outflow port. Compared with the prior art, the utility model has the advantages of practice thrift manpower resources, improve the experiment accuracy, reduce the antibody extravagant, practice thrift the experiment cost.
Description
Technical Field
The utility model belongs to the technical field of molecular biology experiment technique and specifically relates to an automatic western blotting is with antibody incubation box is related to.
Background
Western blotting (Western Blot) is a commonly used molecular biology experimental technique, and aims to transfer target proteins to a PVDF membrane after electrophoretic separation, hybridize with known protein antibodies, and finally perform qualitative and quantitative analysis on the target proteins through a color reaction of an enzyme system. The Western Blot comprises five main steps of electrophoresis, transfer, blocking, hybridization and color development, wherein the blocking and hybridization processes can be called an antibody incubation process together. Generally, in the antibody incubation process, the PVDF membrane is placed in a common rectangular plastic box or a round culture dish for antibody incubation, and placed on a shaking table, and then primary antibody is sequentially added and kept overnight, the membrane is washed three times with TBTS, secondary antibody is added, the membrane is washed three times with TBTS again, and finally color development is performed. This conventional process has some problems: (1) the incubation operation of a rectangular plastic box or a circular culture dish is very inconvenient, cannot be adjusted according to the size of a membrane, and easily causes uneven incubation of an antibody and friction force, so that the result repeatability is poor; (2) the sealing film of the incubation box is not firm because of wetting; (3) an experimenter needs to continuously perform simple mechanical operations such as liquid adding, liquid changing and the like, and a large amount of manpower is consumed; (4) the antibody incubation process is long, and a great deal of time is needed for experimenters; and (5) in the manual operation process, the problems of wrong liquid adding, insufficient antibody recovery, incomplete membrane washing and the like can occur. Therefore, how to complete the Western blot antibody incubation process efficiently and conveniently is a concerned problem and needs to be solved urgently.
The issued patent at present has some designs for some of the problems, such as: patent No. CN 206876712U discloses a design that can adjust the size of the gap by moving the partition plate in the incubation box, and it was preliminarily proposed to solve the problem of mismatch between the membrane and the box. Patent No. CN 205665255U discloses a hydrophobic intermediate partition plate added in an incubation box, wherein the amount of water added is controlled at the lower layer of the partition plate, so that the humidity can be kept, the amount of antibody can be reduced, the volatilization of water can be reduced, and the tightness can be improved.
However, for other problems in the Western Blot antibody incubation process, no patent is provided for a corresponding solution.
SUMMERY OF THE UTILITY MODEL
The utility model aims to overcome the defects of the prior art and provide an automatic antibody incubation box for western blotting.
The purpose of the utility model can be realized through the following technical scheme: the antibody incubation box for the automatic Western blotting method comprises a box cover and a box body, and is characterized in that a liquid storage device is arranged above the box cover, a recovery liquid storage device is arranged below the box body and is arranged in a shell with refrigeration and light-shielding functions, and a plurality of partition plates are arranged in the box body and divide the box body into a plurality of PVDF membrane storage cavities; the top of the box cover is connected with the liquid storage device above the box cover, and the bottom of the box body is connected with the liquid storage device below the box body.
The box cover is provided with a primary antibody adding hole, a TBST adding hole and a secondary antibody adding hole; the reservoir that lid top set up include one anti reservoir an, TBST reservoir an and resist reservoir an, anti reservoir an connect one through the cavity pipeline and resist the interpolation hole, TBST reservoir a connect the TBST interpolation hole through the cavity pipeline, resist reservoir an and connect through the cavity pipeline and resist the interpolation hole.
The primary antibody liquid storage device a is provided with a plurality of, for example 7-10, the TBST liquid storage device a and the secondary antibody liquid storage device a are respectively provided with 1, and each liquid storage device opening is provided with a control valve which is respectively connected with each hollow pipeline. The liquid storage device can realize the functions of avoiding light and the like, and the quantity and the sequence of liquid adding are controlled by opening and closing the valve of the liquid storage device at regular time.
The box cover is provided with three rows of holes, the number of each row of holes is the same as that of the PVDF film storage cavities, the first row of holes are primary anti-adding holes, the second row of holes are TBST adding holes, and the third row of holes are secondary anti-adding holes.
Or, the box cover (1) is provided with a row of holes, the box cover (1) above each PVDF film storage cavity corresponds to one hole, each hole is simultaneously connected with the first-resistance liquid accumulator a (51), the TBST liquid accumulator a (52) and the second-resistance liquid accumulator a (53), the opening and closing of each liquid accumulator are controlled through a valve, and different liquids are input into each PVDF film storage cavity in sequence.
The box cover is made of black smooth plastic, and sealing glue is arranged on the inner side of the box cover.
The outer side of the long shaft of the box body is provided with a scale, the inner side of the long shaft of the box body is provided with a baffle plate with an adjustable position, and the baffle plate (4) is arranged on a slide way in the box body and is positioned by a limiter.
The bottom of the box body is an inclined plane, a partition layer with micropores is arranged above the bottom of the box body, an outflow port is arranged at the tail end of the inclined plane, and liquid in the box body is collected to the outflow port and flows to a recovery liquid storage device at the bottom of the box body through a one-to-many hollow pipe.
The one-to-many hollow pipe is a main pipe connected with the outflow port, and a plurality of parallel branch pipes are branched from the main pipe, each branch pipe is respectively connected with each recovery liquid storage device, and valves opened and closed by a controller are arranged at each recovery liquid storage device and each parallel branch pipe, so that different liquids can be recovered into the corresponding liquid storage devices.
The inclination angle of box body bottom be 3 ~ 10 degrees.
The bottom of the box body is funnel-shaped, and an interlayer with micropores is arranged above the funnel.
The reservoir that the box body bottom set up include one resist reservoir b, TBST reservoir b and resist reservoir b.
Compared with the prior art, the utility model discloses box is hatched to automatic antibody can realize following function: (1) the width of the film channel can be simply and conveniently adjusted; (2) adding a sealing belt of the incubation box to realize tight sealing; (3) automatic timing liquid adding and liquid recovering processes; (4) the outer cover can realize refrigeration and keep the activity of the antibody. Finally, the aims of saving human resources, improving the accuracy of the experiment, reducing the waste of antibodies and saving the experiment cost are achieved.
Drawings
FIG. 1 is a schematic diagram of the overall structure of the antibody incubation cassette for automated Western blotting according to the present invention;
FIG. 2 is a schematic structural view of the upper part of the box cover;
FIG. 3 is a bottom view of the lid;
FIG. 4 is a top view of the cartridge body;
FIG. 5 is a side view of a first cassette construction;
FIG. 6 is a bottom view of the first cassette construction;
FIG. 7 is a side view of a second cassette structure
FIG. 8 is a bottom view of the second cassette construction;
FIG. 9 is a schematic view of a one-in-three hollow pipe structure;
FIG. 10 is a schematic diagram showing the overall structure of an antibody incubation cassette for a second automated Western blotting method;
FIG. 11 is a schematic diagram showing the structure of the upper part of the cassette cover of the antibody incubation cassette for the second automated Western blotting method.
Detailed Description
The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.
Example 1
As shown in fig. 1, an antibody incubation box for an automatic western blotting method comprises a box cover 1 and a box body 2, wherein a liquid storage device is arranged above the box cover 1, a recovery liquid storage device is arranged below the box body 2 and is arranged in a shell 3 with refrigeration and light-shielding functions, a plurality of partition plates 4 are arranged in the box body 2 to divide the box body into a plurality of PVDF film storage cavities 7; the liquid storage device above the box cover 1 is connected with the box cover 1, and the recovery liquid storage device below the box body 2 is connected with the bottom of the box body 2.
As shown in fig. 2 to 3, a first anti-additive hole 11, a TBST additive hole 12 and a second anti-additive hole 13 are formed in the box cover 1; the reservoir that lid 1 top set up include one resist reservoir a51, TBST reservoir a52 and resist reservoir a53, first resist reservoir a51 connect one through the hollow pipeline and resist interpolation hole 11, TBST reservoir a52 connect TBST interpolation hole 12 through the hollow pipeline, resist reservoir a53 and connect through the hollow pipeline and resist interpolation hole 13.
The number of the primary anti liquid accumulators a51 is 7-10, the number of the TBST liquid accumulators a52 and the number of the secondary anti liquid accumulators a53 are 1, and valves for controlling flow are arranged at ports of the liquid accumulators. The liquid storage device can realize the functions of avoiding light and the like, and the quantity and the time sequence of liquid adding are controlled by opening and closing the valve of the liquid storage device at regular time.
As shown in fig. 3, the box cover 1 is provided with three rows of holes, the number of each row of holes is the same as that of the PVDF film storage cavities, the first row of holes are primary anti-additive holes 11, the second row of holes are TBST additive holes 12, and the third row of holes are secondary anti-additive holes 13. The box cover 1 is made of black smooth plastic, and sealing glue is arranged on the inner side of the box cover 1.
As shown in fig. 4, a scale is arranged on the outer side of the long axis of the box body 2, a position-adjustable partition plate 4 is arranged on the inner side of the long axis, the partition plate 4 is arranged on a slideway in the box body and is positioned by a stopper, and the partition plate 4 is fixed by the stopper after being adjusted to a fixed position.
As shown in fig. 5-6, the bottom of box 2 is an inclined plane 21 with an inclination angle of 3-5 degrees, a partition layer a22 with micropores is arranged above, an outlet 23 is arranged at the end of the inclined plane, and the liquid in box 2 is collected to the outlet and flows to a recovery reservoir at the bottom of the box.
The recovery liquid storage device arranged at the bottom of the box body 2 comprises 7-10 primary antibody recovery liquid storage devices b61, a TBST recovery liquid storage device b62 and a secondary antibody recovery liquid storage device b 63. The outflow port 23 is connected to the corresponding reservoir through a three-in-one hollow pipe, wherein an anti-outflow pipe is divided into 7-10 pipes to be reserved to the corresponding reservoir. As shown in fig. 9, the one-in-three hollow pipe is a main pipe 231 connected to the outflow port 23, and a plurality of parallel branch pipes 232 branched from the main pipe 231, each branch pipe 232 is connected to each recovery reservoir (a primary antibody recovery reservoir b61, a TBST recovery reservoir b62, and a secondary antibody recovery reservoir b63), and each recovery reservoir and each parallel branch pipe are provided with a valve 233 opened and closed by a controller, so as to recover different liquids into the corresponding reservoirs.
The upper part of the shell 3 is provided with an upper liquid storage device and a middle box cover 1 of the incubation box, the lower part of the shell is provided with a lower recovery liquid storage device of the incubation box body 2, and the shell 3 has a refrigeration function.
The using method comprises the following steps:
1. opening the housing 3;
2. adjusting the width of each membrane track of the incubation box: the position of the partition plate 4 is adjusted and fixed through the adjusting slide way, so that the size of the PVDF film storage cavity 7-10 is adjusted, and the function of adjusting the width of the film channel is realized;
3. the PVDF membrane is placed in an incubation box (7-10 membranes can be placed, one for each PVDF membrane storage chamber 7), and it is noted that each PVDF membrane storage chamber 7 corresponds to at least 1 primary antibody addition well 11, one TBTS addition well 12 and one secondary antibody addition well 13.
4. Adding sufficient primary antibody, TBTS liquid and secondary antibody into the prepared primary antibody reservoir a51, TBST reservoir a52 and secondary antibody reservoir a53 respectively;
5. setting the opening and closing time, times and sequence of each liquid reservoir valve and the recovery liquid reservoir valve, opening one of the liquid reservoir valves, inputting corresponding liquid into the box body, and incubating the membrane placed in the PVDF membrane storage cavity 7;
6. placing the box body on a shaking table for shaking, opening a valve of a recovery liquid storage device when the incubation time is over, collecting the liquid to an outflow port 23 through an inclined surface 21 at the bottom of the box body 2, and recovering the liquid to the recovery liquid storage device corresponding to the bottom through a hollow pipeline;
7. opening another liquid storage device at the top of the box cover as required, and repeating the step 5-6; and sequentially realizing the processes of primary antibody incubation, TBTS washing, secondary antibody incubation and TBTS washing.
8. After the whole process is finished, a prompt tone can be sent out.
Example 2
As shown in fig. 6-7, the bottom of the box body 2 is funnel-shaped 24, a partition layer b25 with micropores is arranged above the funnel, and the bottom is provided with a liquid outlet 26. The rest is the same as example 1.
Example 3
As shown in fig. 10-11, a row of holes is arranged on the box cover 1, one hole 101 corresponds to the box cover 1 above each PVDF film storage cavity, and each hole 101 is simultaneously connected with the primary antibody liquid reservoir a51, the TBST liquid reservoir a52 and the secondary antibody liquid reservoir a53, the opening and closing of each liquid reservoir is controlled by a valve, and different liquids are sequentially input into each PVDF film storage cavity.
The rest is the same as example 1.
Claims (10)
1. An antibody incubation box for an automatic western blotting method comprises a box cover (1) and a box body (2), and is characterized in that a liquid storage device is arranged above the box cover (1), a recovery liquid storage device is arranged below the box body (2) and is arranged in a shell (3) with refrigeration and light-shielding functions, a plurality of partition plates (4) are arranged in the box body (2) to divide the box body into a plurality of PVDF film storage cavities (7); the liquid storage device above the box cover (1) is connected with the top of the box cover (1), and the liquid storage device below the box body (2) is connected with the bottom of the box body (2).
2. The automatic western blotting antibody incubation box according to claim 1, wherein said box cover (1) is provided with an anti-additive well (11), a TBST additive well (12) and a secondary antibody additive well (13); lid (1) top set up the reservoir include one resist reservoir a (51), TBST reservoir a (52) and resist reservoir a (53), one resist reservoir a (51) connect one through the hollow pipeline and resist interpolation hole (11), TBST reservoir a (52) connect TBST interpolation hole (12) through the hollow pipeline, resist reservoir a (53) connect through the hollow pipeline and resist interpolation hole (13).
3. The automatic western blotting antibody incubation cassette of claim 2, wherein said primary antibody reservoir a (51) is provided in plurality, said TBST reservoir a (52) and secondary antibody reservoir a (53) are each provided in 1, and each reservoir port is provided with a valve for controlling opening and closing, and is connected to each hollow pipe.
4. The automatic western blotting antibody incubation box according to claim 2, wherein the box cover (1) is provided with three rows of holes, the number of each row of holes is the same as the number of the PVDF membrane storage cavities, the first row of holes are primary antibody addition holes (11), the second row of holes are TBST addition holes (12), and the third row of holes are secondary antibody addition holes (13);
or, the box cover (1) is provided with a row of holes, the box cover (1) above each PVDF film storage cavity corresponds to one hole, each hole is simultaneously connected with the first-resistance liquid accumulator a (51), the TBST liquid accumulator a (52) and the second-resistance liquid accumulator a (53), the opening and closing of each liquid accumulator are controlled through a valve, and different liquids are input into each PVDF film storage cavity in sequence.
5. The antibody incubation box for the automatic western blotting method according to claim 1, wherein the box cover (1) is black smooth plastic, and the inner side of the box cover (1) is provided with sealing glue.
6. The automatic western blotting incubation box for antibody according to claim 1, wherein the outside of the long axis of the box body (2) is provided with a scale, the inside is provided with a baffle plate (4) with adjustable position, the baffle plate (4) is arranged on a slide way in the box body and is positioned by a stopper.
7. The automatic western blotting antibody incubation cassette of claim 1, wherein the bottom of the cassette body (2) is an inclined plane (21), and a partition layer a (22) with micropores is provided above the inclined plane, and an outflow port (23) is provided at the end of the inclined plane, the liquid in the cassette body (2) is collected to the outflow port, and flows to a recovery reservoir at the bottom of the cassette body through a divided-by hollow pipe;
the one-to-many hollow pipe is a main pipe (231) connected with the outflow port, and a plurality of parallel branch pipes (232) branched from the main pipe, each branch pipe is respectively connected with each recovery liquid storage device, and valves (233) opened and closed by a controller are arranged at each recovery liquid storage device and each parallel branch pipe.
8. The antibody incubation box for the automatic western blotting method according to claim 7, wherein the inclination angle of the bottom of the box body (2) is 3-10 degrees.
9. The automatic western blotting incubation box according to claim 1, wherein the bottom of the box body (2) is funnel-shaped, and a partition layer b (25) with micropores is arranged above the funnel.
10. The automatic western blotting incubation cassette according to claim 1, wherein the reservoirs provided at the bottom of the cassette body (2) comprise an anti-reservoir b (61), a TBST reservoir b (62), and a secondary antibody reservoir b (63).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201821843301.0U CN209858588U (en) | 2018-11-09 | 2018-11-09 | Antibody incubation box for automatic western blotting method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201821843301.0U CN209858588U (en) | 2018-11-09 | 2018-11-09 | Antibody incubation box for automatic western blotting method |
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| CN209858588U true CN209858588U (en) | 2019-12-27 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109254160A (en) * | 2018-11-09 | 2019-01-22 | 上海交通大学医学院附属第九人民医院 | A kind of automatic Western Blot antibody incubation box |
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- 2018-11-09 CN CN201821843301.0U patent/CN209858588U/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109254160A (en) * | 2018-11-09 | 2019-01-22 | 上海交通大学医学院附属第九人民医院 | A kind of automatic Western Blot antibody incubation box |
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