An automatic Western Blot antibody incubation box

Abstract

The present invention relates to an automatic Western Blot antibody incubation box, comprising a box cover (1) and a box body (2), wherein a liquid reservoir is arranged above the box cover (1), a recovery liquid reservoir is arranged below the box body (2), and the recovery liquid reservoir is placed in a housing (3) with refrigeration and light-shielding functions, and a plurality of partitions (4) are arranged in the box body (2), which divide the box body into a plurality of PVDF membrane storage chambers (7); the liquid reservoir above the box cover (1) is connected to the top of the box cover (1), and the liquid reservoir below the box body (2) is connected to the bottom of the box body (2); the box cover (1) is provided with a primary antibody addition hole, a TBTS addition hole and a secondary antibody addition hole, and the bottom of the box body (2) is provided with an outflow port. Compared with the prior art, the present invention has the advantages of saving human resources, improving experimental accuracy, reducing antibody waste, saving experimental costs, etc.

Description

Technical Field

[0001] The invention relates to the technical field of molecular biology experiments, in particular to an automatic Western Blot antibody incubation box. Background Art

[0002] Western Blot is a commonly used molecular biology experimental technique, the purpose of which is to transfer the target protein to a PVDF membrane after separation by electrophoresis, then hybridize with a known protein antibody, and finally perform qualitative and quantitative analysis of the target protein through the color development reaction of the enzyme system. The main steps of Western Blot include electrophoresis, transfer, blocking, hybridization and color development, among which the blocking and hybridization processes can be collectively referred to as the antibody incubation process. Generally speaking, during the antibody incubation process, the PVDF membrane needs to be placed in a commonly used rectangular plastic box or round culture dish for antibody incubation and placed on a shaker. After that, the primary antibody needs to be added in sequence and overnight, the membrane needs to be washed three times with TBTS, the secondary antibody is added, and the membrane is washed three times with TBTS again, and finally the color is developed. This traditional process has some problems: (1) The rectangular plastic box or round culture dish is very inconvenient to incubate, and it cannot be adjusted according to the size of the membrane, which can easily cause uneven antibody incubation and generate friction, resulting in poor reproducibility of results; (2) The sealing film of the incubation box is not firmly sealed due to moisture; (3) The experimenter needs to constantly perform simple mechanical operations such as adding liquid and changing liquid, which consumes a lot of manpower; (4) The antibody incubation process is long and requires the experimenter to spend a lot of time; (5) During the manual operation, there may be problems such as error in adding liquid, insufficient antibody recovery, and incomplete membrane washing. Therefore, how to complete the Western blot antibody incubation process efficiently and simply is a problem we are concerned about and needs to be solved urgently.

[0003] The currently authorized patents have made some designs for some of these problems. For example, patent number "CN206876712U" has designed a design that can adjust the size of the gap by moving the partition in the incubation box, and preliminarily proposed a solution to the problem of mismatch between the membrane and the box. Patent number "CN 205665255U" has designed a design that adds a hydrophobic middle partition in the incubation box, and controls the amount of water added under the partition to maintain moisture, reduce the amount of antibodies, reduce water volatilization, and improve airtightness.

[0004] However, no patent has yet proposed corresponding solutions to other problems in the Western Blot antibody incubation process. Summary of the invention

[0005] The purpose of the present invention is to provide an automatic Western Blot antibody incubation box in order to overcome the defects of the above-mentioned prior art.

[0006] The purpose of the present invention can be achieved through the following technical scheme: an automatic Western Blot antibody incubation box, comprising a box cover and a box body, characterized in that a liquid reservoir is provided above the box cover, a recovery liquid reservoir is provided below the box body, and is placed in a shell with refrigeration and light-proof functions, and a plurality of partitions are provided in the box body to divide the box body into a plurality of PVDF membrane storage chambers; the liquid reservoir above the box cover is connected to the top of the box cover, and the liquid reservoir below the box body is connected to the bottom of the box body.

[0007] The box cover is provided with a primary antibody addition hole, a TBST addition hole and a secondary antibody addition hole; the liquid reservoirs arranged above the box cover include a primary antibody liquid reservoir a, a TBST liquid reservoir a and a secondary antibody liquid reservoir a, the primary antibody liquid reservoir a is connected to the primary antibody addition hole through a hollow pipe, the TBST liquid reservoir a is connected to the TBST addition hole through a hollow pipe, and the secondary antibody liquid reservoir a is connected to the secondary antibody addition hole through a hollow pipe.

[0008] The primary antibody reservoir a is provided with a plurality of, for example, 7-10, TBST reservoir a and secondary antibody reservoir a are each provided with one, and a control valve is provided at the mouth of each reservoir, respectively connected to each hollow pipe. The reservoir can realize functions such as light protection, and the amount and order of liquid addition are controlled by opening and closing the valve of the reservoir at a fixed time.

[0009] The box cover is provided with three rows of holes, the number of holes in each row is the same as the number of PVDF membrane storage chambers, the first row of holes are primary antibody addition holes, the second row of holes are TBST addition holes, and the third row of holes are secondary antibody addition holes.

[0010] Alternatively, the box cover (1) is provided with a row of holes, each PVDF membrane storage cavity is provided with a corresponding hole on the box cover (1), and each hole is simultaneously connected to the primary antibody reservoir a (51), TBST reservoir a (52) and secondary antibody reservoir a (53), and the opening and closing of each reservoir is controlled by a valve, so that different liquids are successively input into each PVDF membrane storage cavity.

[0011] The box cover is made of black smooth plastic, and sealing glue is arranged on the inner side of the box cover.

[0012] A scale is arranged outside the long axis of the box body, and a position-adjustable partition is arranged inside. The partition (4) is arranged on a slideway inside the box body and is positioned by a limiter.

[0013] The bottom of the box body is an inclined surface, and a partition with micropores is provided on the top, and a flow outlet is provided at the end of the inclined surface. The liquid in the box body is collected at the flow outlet and flows to the recovery liquid storage device at the bottom of the box body through a multi-point hollow tube.

[0014] The one-to-many hollow pipe is a main pipe connected to the outflow outlet, and multiple parallel branch pipes branched from the main pipe, each branch pipe is connected to each recovery liquid reservoir, and each recovery liquid reservoir and each parallel branch pipe is provided with a valve opened and closed by a controller to realize the recovery of different liquids into the corresponding liquid reservoir.

[0015] The inclination angle of the bottom of the box body is 3 to 10 degrees.

[0016] The bottom of the box body is in a funnel shape, and a partition with micropores is arranged above the funnel.

[0017] The liquid reservoirs arranged at the bottom of the box body include a primary antibody liquid reservoir b, a TBST liquid reservoir b and a secondary antibody liquid reservoir b.

[0018] Compared with the prior art, the automatic antibody incubation box of the present invention can achieve the following functions: (1) to achieve simple adjustment of the width of the membrane channel; (2) to add the sealing tape of the incubation box to achieve a tight seal; (3) to automatically add liquid at a fixed time, recover liquid, etc.; (4) the outer cover can achieve refrigeration to maintain the activity of the antibody. Ultimately, the goal of saving human resources, improving experimental accuracy, reducing antibody waste, and saving experimental costs is achieved. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 Schematic diagram of the overall structure of the automatic Western Blot antibody incubation box of the present invention;

[0020] Figure 2 It is a structural schematic diagram of the upper part of the box cover;

[0021] Figure 3 This is a bottom view of the box cover;

[0022] Figure 4 is a top view of the box;

[0023] Figure 5 It is a side view of the first box structure;

[0024] Figure 6 It is a bottom view of the first box structure;

[0025] Figure 7 Side view of the second box structure

[0026] Figure 8 It is a bottom view of the second box structure;

[0027] Fig. 9 This is a schematic diagram of a one-to-three hollow pipe structure;

[0028] Fig.10 It is a schematic diagram of the overall structure of the second automatic Western Blot antibody incubation box;

[0029] Fig.11 Schematic diagram of the structure of the upper part of the lid of the second automatic Western Blot antibody incubation box. DETAILED DESCRIPTION

[0030] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments.

[0031] Example 1

[0032] like Figure 1 As shown, an automatic Western Blot antibody incubation box includes a box cover 1 and a box body 2, wherein a liquid reservoir is provided above the box cover 1, and a recovery liquid reservoir is provided below the box body 2, and is placed in a shell 3 with refrigeration and light-shielding functions, and a plurality of partitions 4 are provided in the box body 2 to divide the box body into a plurality of PVDF membrane storage chambers 7; the liquid reservoir above the box cover 1 is connected to the box cover 1, and the recovery liquid reservoir below the box body 2 is connected to the bottom of the box body 2.

[0033] like Figures 2-3 As shown, the box cover 1 is provided with a primary antibody addition hole 11, a TBST addition hole 12 and a secondary antibody addition hole 13; the liquid reservoirs arranged above the box cover 1 include a primary antibody liquid reservoir a51, a TBST liquid reservoir a52 and a secondary antibody liquid reservoir a53, the primary antibody liquid reservoir a51 is connected to the primary antibody addition hole 11 through a hollow pipe, the TBST liquid reservoir a52 is connected to the TBST addition hole 12 through a hollow pipe, and the secondary antibody liquid reservoir a53 is connected to the secondary antibody addition hole 13 through a hollow pipe.

[0034] The primary antibody reservoir a51 is provided with 7-10, the TBST reservoir a52 and the secondary antibody reservoir a53 are provided with 1 each, and a valve for controlling the flow rate is provided at the mouth of each reservoir. The reservoir can realize functions such as light protection, and the amount and time sequence of liquid addition are controlled by opening and closing the valve of the reservoir at a fixed time.

[0035] like Figure 3 As shown, the box cover 1 is provided with three rows of holes, the number of holes in each row is the same as the number of PVDF membrane storage chambers, the first row of holes is the primary antibody addition hole 11, the second row of holes is the TBST addition hole 12, and the third row of holes is the secondary antibody addition hole 13. The box cover 1 is made of black smooth plastic, and the inner side of the box cover 1 is provided with sealing glue.

[0036] like Figure 4 As shown, a scale is provided on the outer side of the long axis of the box body 2, and an adjustable partition 4 is provided on the inner side. The partition 4 is provided on a slideway in the box body and is positioned by a limiter. The partition 4 is adjusted to a fixed position and then fixed by the limiter.

[0037] like Figure 5-6As shown, the bottom of the box body 2 is an inclined surface 21 with an inclination angle of 3-5 degrees, and a partition layer 22 with micropores is provided on the top, and a flow outlet 23 is provided at the end of the inclined surface. The liquid in the box body 2 is collected at the flow outlet and flows to the recovery liquid storage tank at the bottom of the box body.

[0038] The recovery reservoirs arranged at the bottom of the box body 2 include 7-10 primary antibody recovery reservoirs b61, TBST recovery reservoirs b62 and secondary antibody recovery reservoirs b63. ​​The outflow port 23 is connected to the corresponding reservoirs through a one-to-three hollow tube, wherein the primary antibody outflow tube is further divided into 7-10 and retained in the corresponding reservoirs. The one-to-three hollow tubes are as follows Fig. 9 As shown, there is a main pipe 231 connected to the outflow port 23, and a plurality of parallel branch pipes 232 branching from the main pipe 231, each branch pipe 232 is respectively connected to each recovery reservoir (primary antibody recovery reservoir b61, TBST recovery reservoir b62 and secondary antibody recovery reservoir b63), and each recovery reservoir and each parallel branch pipe is provided with a valve 233 which is opened and closed by a controller to realize the recovery of different liquids into the corresponding reservoir.

[0039] The upper part of the shell 3 is equipped with an upper liquid reservoir + a box cover 1 of the middle incubation box, and the lower part of the shell is equipped with an incubation box body 2 + a lower recovery liquid reservoir. The shell 3 has a refrigeration function.

[0040] Here’s how to use it:

[0041] 1. Open the housing 3;

[0042] 2. Adjust the width of each membrane channel of the incubation box: adjust and fix the position of the partition 4 by adjusting the slideway, thereby adjusting the size of the PVDF membrane storage cavity 7-10, thereby achieving the function of adjusting the width of the membrane channel;

[0043] 3. Place the PVDF membrane into the incubation box (7-10 membranes can be placed, one in each PVDF membrane storage chamber 7). Note that each PVDF membrane storage chamber 7 corresponds to at least one primary antibody addition hole 11, one TBTS addition hole 12 and one secondary antibody addition hole 13.

[0044] 4. Add sufficient amount of primary antibody, TBTS solution and secondary antibody to the designated primary antibody reservoir a51, TBST reservoir a52 and secondary antibody reservoir a53 respectively;

[0045] 5. Set the opening and closing time, frequency and sequence of each reservoir valve and the recovery reservoir valve, open the valve of one of the reservoirs, input the corresponding liquid into the box body, and incubate the membrane placed in the PVDF membrane storage chamber 7;

[0046] 6. Place the incubator on a shaker and shake. When the incubation time is over, the valve of the recovery reservoir is opened, and the liquid flows through the inclined surface 21 at the bottom of the box body 2, collects at the outflow port 23, and is recovered to the corresponding recovery reservoir at the bottom through the hollow pipe;

[0047] 7. Open another reservoir on the top of the box lid as needed and repeat steps 5-6; carry out the process of primary antibody incubation - TBTS washing - secondary antibody incubation - TBTS washing in sequence.

[0048] 8. A prompt sound will be issued after the whole process is completed.

[0049] Example 2

[0050] like Figure 6-7 As shown, the bottom of the box body 2 is in a funnel shape 24, and a partition layer 25 with micropores is provided above the funnel, and a liquid outflow outlet 26 is provided at the bottom. The rest is the same as in the first embodiment.

[0051] Example 3

[0052] like Figure 10-11 As shown, a row of holes is provided on the box cover 1, and each PVDF membrane storage cavity above the box cover 1 corresponds to a hole 101, and each hole 101 is simultaneously connected to the primary antibody reservoir a51, TBST reservoir a52 and secondary antibody reservoir a53. The opening and closing of each reservoir is controlled by a valve, and different liquids are successively input into each PVDF membrane storage cavity.

[0053] The rest is the same as in Example 1.

Claims

1. An automatic Western Blot antibody incubation box, comprising a box cover (1) and a box body (2), characterized in that: A liquid reservoir is provided above the box cover (1), and a recovery liquid reservoir is provided below the box body (2), and is placed in a housing (3) with refrigeration and light-shielding functions. A plurality of partitions (4) are provided inside the box body (2), which divide the box body into a plurality of PVDF membrane storage chambers (7); the liquid reservoir above the box cover (1) is connected to the top of the box cover (1), and the liquid reservoir below the box body (2) is connected to the bottom of the box body (2); The box cover (1) is provided with a primary antibody addition hole (11), a TBST addition hole (12) and a secondary antibody addition hole (13); the liquid reservoirs arranged above the box cover (1) include a primary antibody liquid reservoir a (51), a TBST liquid reservoir a (52) and a secondary antibody liquid reservoir a (53); the primary antibody liquid reservoir a (51) is connected to the primary antibody addition hole (11) via a hollow pipe, the TBST liquid reservoir a (52) is connected to the TBST addition hole (12) via a hollow pipe, and the secondary antibody liquid reservoir a (53) is connected to the secondary antibody addition hole (13) via a hollow pipe; there are a plurality of primary antibody liquid reservoirs a (51), and there are one TBST liquid reservoir a (52) and one secondary antibody liquid reservoir a (53); a valve for controlling opening and closing is provided at the mouth of each liquid reservoir, and each of the valves is connected to each hollow pipe; The box body (2) is provided with a scale on the outside of the long axis, and a position-adjustable partition (4) is provided on the inside; the partition (4) is provided on a slideway in the box body and is positioned by a limiter; Each PVDF membrane storage chamber (7) corresponds to at least one primary antibody addition hole (11), one TBST addition hole (12) and one secondary antibody addition hole (13); The bottom of the box body (2) is an inclined surface (21), and a partition layer (22) with micropores is provided above the inclined surface. At the same time, a flow outlet (23) is provided at the end of the inclined surface. The liquid in the box body (2) is collected at the flow outlet and flows to the recovery liquid storage device at the bottom of the box body through a multi-point hollow tube. The one-to-many hollow pipe is a main pipe (231) connected to the outflow outlet, and a plurality of parallel branch pipes (232) branching from the main pipe, each branch pipe is connected to a recovery liquid reservoir, and each recovery liquid reservoir and each parallel branch pipe is provided with a valve (233) opened and closed by a controller.

2. An automatic Western Blot antibody incubation box according to claim 1, characterized in that: The box cover (1) is provided with three rows of holes, the number of holes in each row is the same as the number of PVDF membrane storage chambers, 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); Alternatively, the box cover (1) is provided with a row of holes, each PVDF membrane storage cavity is provided with a corresponding hole on the box cover (1), and each hole is simultaneously connected to the primary antibody reservoir a (51), TBST reservoir a (52) and secondary antibody reservoir a (53), and the opening and closing of each reservoir is controlled by a valve, so that different liquids are successively input into each PVDF membrane storage cavity.

3. An automatic Western Blot antibody incubation box according to claim 1, characterized in that: The box cover (1) is made of black smooth plastic, and a sealing glue is provided on the inner side of the box cover (1).

4. An automatic Western Blot antibody incubation box according to claim 1, characterized in that: The inclination angle of the bottom of the box body (2) is 3 to 10 degrees.

5. An automatic Western Blot antibody incubation box according to claim 1, characterized in that: The bottom of the box body (2) is funnel-shaped, and a partition layer (25) with micropores is provided above the funnel.

6. An automatic Western Blot antibody incubation box according to claim 1, characterized in that: The liquid reservoirs arranged at the bottom of the box body (2) include a primary antibody liquid reservoir b (61), a TBST liquid reservoir b (62) and a secondary antibody liquid reservoir b (63).

Images