CN111596049A - Novel antibody incubation device and use method thereof - Google Patents
Novel antibody incubation device and use method thereof Download PDFInfo
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- CN111596049A CN111596049A CN202010648370.1A CN202010648370A CN111596049A CN 111596049 A CN111596049 A CN 111596049A CN 202010648370 A CN202010648370 A CN 202010648370A CN 111596049 A CN111596049 A CN 111596049A
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- 238000011534 incubation Methods 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 51
- 239000012528 membrane Substances 0.000 claims description 42
- 239000003085 diluting agent Substances 0.000 claims description 17
- 239000012154 double-distilled water Substances 0.000 claims description 17
- 238000009396 hybridization Methods 0.000 claims description 17
- 229920003023 plastic Polymers 0.000 claims description 14
- 239000000463 material Substances 0.000 claims description 10
- 239000004575 stone Substances 0.000 claims description 10
- 239000002033 PVDF binder Substances 0.000 claims description 9
- 229920002981 polyvinylidene fluoride Polymers 0.000 claims description 9
- 238000007789 sealing Methods 0.000 claims description 9
- 230000000694 effects Effects 0.000 claims description 5
- 230000005484 gravity Effects 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 238000005086 pumping Methods 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 2
- 238000002474 experimental method Methods 0.000 abstract description 2
- 239000004033 plastic Substances 0.000 description 6
- 239000002699 waste material Substances 0.000 description 5
- 238000004806 packaging method and process Methods 0.000 description 4
- 208000033641 Ring chromosome 5 syndrome Diseases 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- ATJFFYVFTNAWJD-UHFFFAOYSA-N Tin Chemical compound [Sn] ATJFFYVFTNAWJD-UHFFFAOYSA-N 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000012447 hatching Effects 0.000 description 1
- 238000010030 laminating Methods 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 239000012785 packaging film Substances 0.000 description 1
- 229920006280 packaging film Polymers 0.000 description 1
- 239000002985 plastic film Substances 0.000 description 1
- 229920006255 plastic film Polymers 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
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Abstract
The invention belongs to the technical field of reaction kits, and particularly relates to a novel antibody incubation device which comprises a box cover, a box body and a wax block, wherein the box cover is in threaded connection with the upper end of the box body, the wax block is arranged in the box body, and the diameter of the wax block is 0.3-0.7cm smaller than the inner diameter of the box body; the invention has simple integral structure, convenient operation, less antibody demand, high incubation efficiency and repeated use, and provides guarantee for the high-efficiency and smooth experiment.
Description
Technical Field
The invention belongs to the technical field of antibody incubation devices, and particularly relates to a novel antibody incubation device and a using method thereof.
Background
When antibody incubation is performed in an experiment, the traditional antibody incubation method has a plurality of problems:
(1) placing the membrane and the antibody in a small box, and incubating in a shaking table, wherein the method needs a large amount of antibody each time, is difficult to incubate multiple antibodies at the same time, and is incomplete in antibody recovery;
(2) the membrane and the antibody are placed in a small bag made of a disposable plastic packaging membrane, the plastic packaging membrane can be sealed only by using a sealing machine, but the plastic packaging membrane is wetted by liquid and is difficult to be sealed by the sealing machine, so that the antibody diluent is easy to leak out of the plastic packaging membrane, and the antibody waste is caused; secondly, air bubbles in the plastic film cannot be sufficiently excluded when the antibody incubation is performed, and the presence of air bubbles makes the antibody not sufficiently contact with the film, thereby affecting the experimental results. The method causes great waste of time and antibody in sample adding and antibody recovery, and causes resource waste and environmental pollution due to the use of a disposable plastic packaging film;
(3) when carrying out the antibody and hatching, often need carry out the shading operation, but lack supporting shading instrument, need materials such as laboratory staff with tinfoil to carry out the parcel operation, make the antibody landing easily, cause the waste of antibody, and the parcel operation is wasted time and energy, and the operation is inconvenient.
Therefore, it is necessary to provide an incubation cassette and a method of using the same, which is simple to operate, stable in effect and cost-effective.
Disclosure of Invention
The invention provides a novel antibody incubation device and a use method thereof aiming at the problems.
In order to achieve the purpose, the invention adopts the following technical scheme:
the utility model provides a novel antibody is hatched device, includes lid, box body and wax stone, lid threaded connection is in the upper end of box body, the wax stone sets up in the box body, the diameter ratio of wax stone the internal diameter of box body is little 0.3-0.7 cm.
Further, a shading device is arranged above the box cover and consists of a rotating shaft arranged horizontally and two semicircular shading plates arranged on the rotating shaft, the two semicircular shading plates are symmetrically arranged about the axis of the rotating shaft and can rotate around the rotating shaft, a plug-in column is fixedly connected to the middle of the rotating shaft, and a socket used for placing the plug-in column is arranged in the middle of the upper surface of the box cover.
Still further, the up end of box body is provided with the sealing washer, so that the lid prevents liquid seepage or volatilizees with the box body laminating closely.
Furthermore, a cross-shaped groove is formed in the bottom of the box body.
Still further, be provided with the inlet opening in the side middle part of box body the side bottom of box body is provided with the apopore, the apopore sets up the arbitrary one end at the cross recess, so that the apopore is swift smooth when the water is gone out be provided with No. one hole stopper and No. two hole stoppers on inlet opening and the apopore respectively, No. one hole stopper and No. two hole stoppers are the toper hole stopper.
Furthermore, the diameter of the water inlet hole is smaller than that of the water outlet hole.
Furthermore, a connecting band is fixedly connected to the first hole plug and the second hole plug, and the other end of the connecting band is fixedly connected with the outer surface of the box body.
Furthermore, the box cover is made of transparent plastic materials, and the box body is made of non-transparent plastic materials.
The invention also provides a use method of the novel antibody incubation device, which comprises the following steps:
1) injecting double-distilled water pre-cooled in a refrigerator at 4 ℃ into the cross groove at the bottom of the box body through the water inlet hole until the double-distilled water overflows from the cross groove by 0.1-0.3 cm;
2) placing the wax block at the bottom of the box body, checking whether the wax block horizontally floats in the box body, and if the density of the wax block is uneven and the center of gravity is unbalanced, so that the wax block cannot horizontally float in the box body, replacing the wax block until the wax block can horizontally float in the box body;
3) pumping the double distilled water in the groove through the water outlet hole to prevent the wax block from moving in the operation process;
4) dripping 4-8 mul of antibody diluent at the expected reaction position on the wax block, and leading the reaction surface of the hybrid membrane to face the wax block, wherein the antibody diluent is diffused and distributed on the reaction surface of the hybrid membrane due to siphon effect;
5) slightly moving the lower hybridization membrane by using tweezers to make the lower hybridization membrane more fully contacted with the antibody diluent; dripping 1-3 mul of antibody diluent on the back of the hybrid membrane to prevent the hybrid membrane from drying and influencing subsequent reaction;
6) repeating the step 1), injecting double distilled water pre-cooled in a refrigerator at 4 ℃ into the bottom of the box body to level the wax block, so as to ensure the level of the hybrid membrane in the whole antibody incubation process and prevent the occurrence of uneven incubation;
7) covering the box cover on the box body, and locking the box cover in a threaded manner;
8) inserting a shading device on the inserting opening, unfolding the two semicircular shading plates, and shading and protecting the antibody in the box body;
9) placing the operated antibody incubation device in an environment of 4 ℃ for hybridization reaction;
10) during the hybridization process, one of the semicircular light shielding plates can be closed to observe the condition of the antibody.
Further, the hybridization membrane in the step 4) is a PVDF membrane or an NC membrane.
Compared with the prior art, the invention has the following advantages:
1. the box cover and the box body are in threaded connection, so that the box cover and the box body are prevented from falling off from each other when the box is taken;
2. the box body is provided with the sealing ring, so that the box cover is tightly attached to the box body, and liquid is prevented from leaking or volatilizing;
3. the wax block has the moisturizing function, and provides a moist environment for the antibody;
4. the wax block has smooth surface, and can prevent the incubated hybridization membrane from being damaged;
5. the wax block is not dissolved in water, and the antibody incubation operation can be performed on the surface of the wax block, so that the recovery of the antibody after the incubation is finished is facilitated.
6. The wax block can float on a small amount of water, so that the wax block is kept horizontal, the level of a hybridization membrane in the whole antibody incubation process is ensured, and the phenomenon of uneven incubation is prevented;
7. the shading device is arranged on the box cover in an inserted mode, is inserted when shading is needed and is taken down when shading is not needed, and different requirements of antibody incubation are met;
8. the shading device comprises the rotating shaft and the two turnable semicircular shading plates, so that experimenters can conveniently combine one semicircular shading plate to observe the incubation condition of the antibody at any time in the hybridization process;
9. according to the invention, the cross-shaped groove is formed in the bottom of the box body, so that the water storage capacity is increased, the wax block cannot incline due to excessive water at the bottom of the box, the humidity in the box is kept, and the volatilization of an over-dried antibody is prevented;
10. according to the invention, the water outlet hole is arranged at any end of the cross-shaped groove, so that the water outlet hole is rapid and smooth when water is discharged;
11. according to the invention, the hole plugs are fixedly connected with the connecting bands, so that the hole plugs are prevented from being lost in the practical process;
12. according to the invention, the box cover and the box body are made of plastic materials, so that the cleaning by experimenters is facilitated, and the waste of resources caused by damage is reduced.
Drawings
FIG. 1 is a schematic structural view of the present invention;
FIG. 2 is a schematic structural view of the lid of the present invention;
FIG. 3 is a schematic view of the structure of the shading device of the present invention;
FIG. 4 is a schematic diagram of the structure of the cross-shaped groove of the present invention;
FIG. 5 is a schematic structural view of the external thread, the water inlet hole, the water outlet hole, the sealing ring, the first hole plug and the second hole plug of the present invention;
FIG. 6 is a schematic view of the location structure of the wax block of the present invention;
in the drawing, a box cover-1, a box body-2, a wax block-3, a socket-4, a sealing ring-5, a semicircular light screen-6, a rotating shaft-7, a plug column-8, a cross groove-9, a connecting belt-10, a water inlet hole-11, a water outlet hole-12, a first hole plug-13 and a second hole plug-14 are arranged.
Detailed Description
In order to further illustrate the technical solution of the present invention, the present invention is further illustrated by the following examples.
Example 1
As shown in fig. 1 to 6, a novel antibody incubation device comprises a box cover 1, a box body 2 and a wax block 3, wherein the box cover 1 is in threaded connection with the upper end of the box body 2, the box cover 1 is made of transparent plastic material, the box body 2 is made of non-transparent plastic material, a shading device is arranged above the box cover 1, the shading device is composed of a rotating shaft 7 arranged horizontally and two semicircular shading plates 6 arranged on the rotating shaft 7, the two semicircular shading plates 6 are symmetrically arranged about the axis of the rotating shaft 7, the two semicircular shading plates 6 can rotate around the rotating shaft 7, a plug-in post 8 is fixedly connected to the middle of the rotating shaft 7, a socket 4 for placing the plug-in post 8 is arranged in the middle of the upper surface of the box cover 1, a sealing ring 5 is arranged on the upper end surface of the box body 2, so that the box cover 1 and the box body 2 are tightly, the bottom of the box body 2 is provided with a cross groove 9, the middle part of the side surface of the box body 2 is provided with a water inlet 11, the bottom of the side surface of the box body 2 is provided with a water outlet 12, the water outlet 12 is arranged at any end of the cross groove 9, so that the water outlet 12 can rapidly and smoothly discharge water, the diameter of the water inlet 11 is smaller than that of the water outlet 12, the water inlet 11 and the water outlet 12 are respectively provided with a first hole plug 13 and a second hole plug 14, the first hole plug 13 and the second hole plug 14 are both conical hole plugs, the minimum diameter of the first hole plug 13 is equal to that of the water inlet 11, the minimum diameter of the second hole plug 14 is equal to that of the water outlet 12, the maximum diameter of the first hole plug 13 is larger than that of the water inlet 11, and the maximum diameter of the second hole plug 14 is larger than that of the water outlet 12, equal fixedly connected with connecting band 10 on a hole stopper 13 and No. two hole stoppers 14, the other end of connecting band 10 with the surface fixed connection of box body 2, wax stone 3 sets up in the box body 2, the diameter ratio of wax stone 3 the internal diameter of box body 2 is little by 0.3-0.7 cm.
A use method of the novel antibody incubation device comprises the following steps:
1) injecting double-distilled water pre-cooled in a refrigerator at 4 ℃ into a cross groove 9 at the bottom of the box body 2 through a water inlet hole 11 until the double-distilled water overflows from the cross groove by 0.1-0.3 cm;
2) placing the wax block 3 at the bottom of the box body 2, checking whether the wax block 3 horizontally floats in the box body 2, and if the wax block 3 is uneven in density and unbalanced in center of gravity, so that the wax block 3 cannot horizontally float in the box body 2, replacing the wax block 3 until the wax block can horizontally float in the box body 2;
3) pumping the double distilled water in the groove 9 through the water outlet hole 12 to prevent the wax block from moving in the operation process;
4) dripping 4-8 mul of antibody diluent at the expected reaction position on the wax block 3, and leading the reaction surface of the PVDF membrane to face the wax block, wherein the antibody diluent is diffused and distributed on the reaction surface of the PVDF membrane due to siphon effect;
5) slightly moving the lower PVDF membrane by using forceps to make the PVDF membrane more fully contacted with the antibody diluent; dripping 1-3 mul of antibody diluent on the back of the PVDF membrane to prevent the PVDF membrane from being dry at the edges and influencing the subsequent reaction;
6) repeating the step 1), injecting double distilled water pre-cooled in a refrigerator at 4 ℃ into the bottom of the box body 2 to level the wax block 3, so as to ensure the level of the PVDF membrane in the whole antibody incubation process and prevent the occurrence of uneven incubation;
7) the box cover 1 is covered on the box body 2 and locked by screw threads;
8) inserting a shading device on the socket 4, unfolding the two semicircular shading plates 6, and shading and protecting the antibodies in the box body;
9) placing the operated antibody incubation device in an environment of 4 ℃ for hybridization reaction;
10) during the hybridization process, one of the semicircular light shielding plates 6 can be closed to observe the condition of the antibody.
Example 2
As shown in fig. 1 to 6, a novel antibody incubation device comprises a box cover 1, a box body 2 and a wax block 3, wherein the box cover 1 is in threaded connection with the upper end of the box body 2, the box cover 1 is made of transparent plastic material, the box body 2 is made of non-transparent plastic material, a shading device is arranged above the box cover 1, the shading device is composed of a rotating shaft 7 arranged horizontally and two semicircular shading plates 6 arranged on the rotating shaft 7, the two semicircular shading plates 6 are symmetrically arranged about the axis of the rotating shaft 7, the two semicircular shading plates 6 can rotate around the rotating shaft 7, a plug-in post 8 is fixedly connected to the middle of the rotating shaft 7, a socket 4 for placing the plug-in post 8 is arranged in the middle of the upper surface of the box cover 1, a sealing ring 5 is arranged on the upper end surface of the box body 2, so that the box cover 1 and the box body 2 are tightly, the bottom of the box body 2 is provided with a cross groove 9, the middle part of the side surface of the box body 2 is provided with a water inlet 11, the bottom of the side surface of the box body 2 is provided with a water outlet 12, the water outlet 12 is arranged at any end of the cross groove 9, so that the water outlet 12 can rapidly and smoothly discharge water, the diameter of the water inlet 11 is smaller than that of the water outlet 12, the water inlet 11 and the water outlet 12 are respectively provided with a first hole plug 13 and a second hole plug 14, the first hole plug 13 and the second hole plug 14 are both conical hole plugs, the minimum diameter of the first hole plug 13 is equal to that of the water inlet 11, the minimum diameter of the second hole plug 14 is equal to that of the water outlet 12, the maximum diameter of the first hole plug 13 is larger than that of the water inlet 11, and the maximum diameter of the second hole plug 14 is larger than that of the water outlet 12, equal fixedly connected with connecting band 10 on a hole stopper 13 and No. two hole stoppers 14, the other end of connecting band 10 with the surface fixed connection of box body 2, wax stone 3 sets up in the box body 2, the diameter ratio of wax stone 3 the internal diameter of box body 2 is little by 0.3-0.7 cm.
A use method of the novel antibody incubation device comprises the following steps:
1) injecting double-distilled water pre-cooled in a refrigerator at 4 ℃ into a cross groove 9 at the bottom of the box body 2 through a water inlet hole 11 until the double-distilled water overflows from the cross groove by 0.1-0.3 cm;
2) placing the wax block 3 at the bottom of the box body 2, checking whether the wax block 3 horizontally floats in the box body 2, and if the wax block 3 is uneven in density and unbalanced in center of gravity, so that the wax block 3 cannot horizontally float in the box body 2, replacing the wax block 3 until the wax block can horizontally float in the box body 2;
3) pumping the double distilled water in the groove 9 through the water outlet hole 12 to prevent the wax block from moving in the operation process;
4) dripping 4-8 mul of antibody diluent at the expected reaction position on the wax block 3, and leading the NC membrane reaction surface to face the wax block, wherein the antibody diluent is diffused and distributed on the NC membrane reaction surface due to siphon effect;
5) slightly moving the lower NC membrane by using tweezers to ensure that the lower NC membrane is more fully contacted with the antibody diluent; dripping 1-3 mul of antibody diluent on the back of the NC membrane to prevent the NC membrane from being dry and influencing subsequent reaction;
6) repeating the step 1), injecting double distilled water pre-cooled in a refrigerator at 4 ℃ into the bottom of the box body 2 to level the wax block 3, so as to ensure the level of the NC membrane in the whole antibody incubation process and prevent the uneven incubation phenomenon;
7) the box cover 1 is covered on the box body 2 and locked by screw threads;
8) inserting a shading device on the socket 4, unfolding the two semicircular shading plates 6, and shading and protecting the antibodies in the box body;
9) placing the operated antibody incubation device in an environment of 4 ℃ for hybridization reaction;
10) during the hybridization process, one of the semicircular light shielding plates 6 can be closed to observe the condition of the antibody.
While there have been shown and described what are at present considered to be the essential features and advantages of the invention, it will be apparent to those skilled in the art that the invention is not limited to the details of the foregoing exemplary embodiments, but is capable of other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (10)
1. A novel antibody incubation device is characterized in that: including lid (1), box body (2) and wax stone (3), lid (1) threaded connection be in the upper end of box body (2), wax stone (3) set up in box body (2), the diameter ratio of wax stone (3) the internal diameter of box body (2) is little by 0.3-0.7 cm.
2. A novel antibody incubation device according to claim 1, characterized in that: be provided with shade above lid (1), shade comprises axis of rotation (7) that the level set up and two semi-circular light screen (6) of setting on axis of rotation (7), and two semi-circular light screen (6) set up about axis of rotation (7) axis symmetry, and two semi-circular light screen (6) all can revolute axis of rotation (7) and rotate the middle part fixedly connected with plug column (8) of axis of rotation (7) the upper surface middle part of lid (1) is provided with socket (4) that are used for placing plug column (8).
3. A novel antibody incubation device according to claim 1, characterized in that: the upper end face of the box body (2) is provided with a sealing ring (5) so that the box cover (1) is tightly attached to the box body (2) to prevent liquid leakage or volatilization.
4. A novel antibody incubation device according to claim 1, characterized in that: the bottom of the box body (2) is provided with a cross-shaped groove (9).
5. A novel antibody incubation device according to claim 1, characterized in that: the side middle part of box body (2) is provided with inlet opening (11) the side bottom of box body (2) is provided with apopore (12), apopore (12) set up the arbitrary one end in cross recess (9), so that apopore (12) are swift smooth when going out water be provided with No. one hole stopper (13) and No. two hole stoppers (14) on inlet opening (11) and apopore (12) respectively, No. one hole stopper (13) and No. two hole stoppers (14) are the toper hole stopper.
6. A novel antibody incubation device according to claim 5, characterized in that: the diameter of the water inlet hole (11) is smaller than that of the water outlet hole (12).
7. A novel antibody incubation device according to claim 5, characterized in that: equal fixedly connected with connecting band (10) on a hole stopper (13) and No. two hole stoppers (14), the other end of connecting band (10) with the surface fixed connection of box body (2).
8. A novel antibody incubation device according to claim 1, characterized in that: the box cover (1) is made of transparent plastic materials, and the box body (2) is made of non-transparent plastic materials.
9. A method of using the novel antibody incubation device of claim 1, comprising the steps of:
1) injecting double-distilled water pre-cooled in a refrigerator at 4 ℃ into a cross groove (9) at the bottom of the box body (2) through a water inlet hole (11), and adding the double-distilled water until the double-distilled water overflows from the cross groove by 0.1-0.3 cm;
2) the wax block (3) is placed at the bottom of the box body (2), whether the wax block (3) horizontally floats in the box body (2) is checked, if the density of the wax block (3) is not uniform and the gravity center is not balanced, the wax block (3) cannot horizontally float in the box body (2), the wax block (3) is replaced until the wax block can horizontally float in the box body (2);
3) pumping the double distilled water in the groove (9) through the water outlet hole (12) to prevent the wax block from moving in the operation process;
4) 4-8 mul of antibody diluent is dripped at the expected reaction position on the wax block (3), the reaction surface of the hybrid membrane faces to the wax block, and the antibody diluent is diffused and distributed on the reaction surface of the hybrid membrane due to siphon effect;
5) slightly moving the lower hybridization membrane by using tweezers to make the lower hybridization membrane more fully contacted with the antibody diluent; dripping 1-3 mul of antibody diluent on the back of the hybrid membrane to prevent the hybrid membrane from drying and influencing subsequent reaction;
6) repeating the step 1), injecting double distilled water pre-cooled in a refrigerator at 4 ℃ into the bottom of the box body (2) to level the wax block (3), so as to ensure the level of the hybrid membrane in the whole antibody incubation process and prevent the occurrence of uneven incubation;
7) the box cover (1) is covered on the box body (2), and the box body is locked by screw threads;
8) the shading device is inserted into the socket (4), and the two semicircular shading plates (6) are unfolded to shade and protect the antibodies in the box body;
9) placing the operated antibody incubation device in an environment of 4 ℃ for hybridization reaction;
10) during the hybridization process, one of the semicircular light shielding plates (6) can be closed to observe the condition of the antibody.
10. The use method of the novel antibody incubation device according to claim 9, characterized in that: the hybridization membrane in the step 4) is a PVDF membrane or an NC membrane.
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| CN202010648370.1A CN111596049A (en) | 2020-07-07 | 2020-07-07 | Novel antibody incubation device and use method thereof |
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| CN202010648370.1A CN111596049A (en) | 2020-07-07 | 2020-07-07 | Novel antibody incubation device and use method thereof |
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