CN116500275A - Laboratory protein development auxiliary device - Google Patents
Laboratory protein development auxiliary device Download PDFInfo
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- CN116500275A CN116500275A CN202310436906.7A CN202310436906A CN116500275A CN 116500275 A CN116500275 A CN 116500275A CN 202310436906 A CN202310436906 A CN 202310436906A CN 116500275 A CN116500275 A CN 116500275A
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- protein
- development
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- film
- baffle
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- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 125
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 125
- 238000011161 development Methods 0.000 title claims abstract description 59
- 238000000034 method Methods 0.000 claims abstract description 22
- 239000007788 liquid Substances 0.000 claims description 6
- 239000012474 protein marker Substances 0.000 claims description 6
- 239000003550 marker Substances 0.000 claims description 5
- 230000002452 interceptive effect Effects 0.000 claims 1
- 239000012528 membrane Substances 0.000 description 5
- 238000003384 imaging method Methods 0.000 description 3
- 238000001179 sorption measurement Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000007306 turnover Effects 0.000 description 1
Classifications
-
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Immunology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Peptides Or Proteins (AREA)
Abstract
The application relates to the field of biological development, in particular to a laboratory protein development auxiliary device. The application provides a protein development auxiliary device, including movable support and the fixed bolster of connection, the movable support includes framework portion and cavity, is equipped with a plurality of baffles on the movable support, the baffle is followed framework portion slides, the bottom of fixed bolster is equipped with the magnetic stripe. According to the method and the device, the purpose that the target strips with a plurality of interference strips are accurately positioned by adjusting the positions of the baffle plates under the condition that the film carrying the protein is not cut can be achieved, and the purpose that the color development result of the target protein is accurately and clearly obtained is achieved.
Description
Technical Field
The application relates to the field of biological development, in particular to a laboratory protein development auxiliary device.
Background
Western immunoblotting (Western Blot) is a protein analysis method commonly used in basic research and clinical laboratories, and chemiluminescence development is the main way to obtain the band of interest. Since it is often necessary to detect multiple target proteins in the same sample in an experiment, three main methods are: (1) According to the quantity of target proteins to be detected, carrying out electrophoresis, membrane transfer, antibody incubation and development on the same sample for corresponding times to obtain the target protein; (2) Cutting the membrane carrying the protein according to the molecular weight of the target protein, and then incubating and developing the antibodies; (3) And (3) stripping the same film for multiple times by using an antibody stripping solution according to the number of target proteins, and incubating different antibodies for development. However, all three methods have drawbacks: firstly, the first method has great influence on experimental results by egg sample loading and repeated stability of the whole operation, and is not feasible for few samples; the second method has higher requirements on the cutting film, is not applicable to proteins with adjacent molecular weights, and is not in line with the requirements of high-quality magazines on data because the whole film cannot be displayed in the obtained data due to the fact that the whole film is cut; the third method has a drawback in that although the antibody stripping solution can remove the bound antibody to some extent, it is destructive to the membrane itself on the one hand, and the unevenness due to the stripping also occurs in some cases, and the real situation of the protein content cannot be obtained; on the other hand, some antibodies are often difficult to strip off, and development of new antibodies is greatly affected. The more general problem is that when there are many nonspecific bands of an antibody for detecting a target protein, the ideal target band information cannot be obtained only by changing the antibody or cutting the membrane, but none is the best method to solve the problem.
The chemiluminescent imager used for protein development at present can develop the region of interest through software, but the problems cannot be effectively solved. There is therefore a need for a device that does not cut the protein-loaded film and that can accurately develop the desired band region during development for use with a chemiluminescent imager.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, in order to solve the technical problem that it is difficult to accurately develop a target strip area in the prior art, an object of the present application is to provide a laboratory protein development auxiliary device, which is used with a chemiluminescent imager, and can accurately develop a target strip area under the condition that a protein-carrying film with an intact protein marker is developed.
To achieve the above object and other related objects, an aspect of the present application provides a protein development auxiliary device, including movable support and the fixed bolster of connection, the movable support includes framework portion and cavity, be equipped with a plurality of baffles on the movable support, the baffle is followed framework portion slides, the bottom of fixed bolster is equipped with the magnetic stripe.
In another aspect the present application provides the use of the aforementioned protein development aid in protein development.
In another aspect, the present application provides a protein development method, which adopts the above-mentioned protein development auxiliary device, and includes the following steps:
1) Placing the protein film in a console of a development imager such that the protein film is positioned in the cavity;
2) Turning the movable bracket relative to the fixed bracket so that the movable bracket does not shade the protein film, and collecting images to obtain a white light image carrying the protein film;
3) Putting down the movable bracket to cover the protein film, sliding and adjusting the baffle according to the position of a marker where the molecular weight of the target protein is located, so that the baffle just exposes the position where the target protein is located, and then adding chemiluminescent liquid to collect chemiluminescent images to obtain a target protein strip chart;
4) Fitting the white light map carrying the protein film obtained in the step 2) and the target protein strip map obtained in the step 3) to obtain a target image.
Compared with the prior art, the beneficial effects of this application are:
1. according to the method and the device, accurate development of a plurality of target strip areas can be achieved by adjusting the positions of the baffles under the condition that the protein-loaded film is not cut.
2. When the used antibody has nonspecific stripes, only the target stripe area is exposed by controlling the baffle distance, and the target stripe color development result is accurately obtained.
3. The device is suitable for most chemiluminescent imagers of different brands, is fixed on the operating platform of the chemiluminescent imagers through magnetic stripe adsorption, does not cause any damage to the operating platform, and is easy to install and detach.
Drawings
Fig. 1 is a top view of the present application.
Fig. 2 is a right side view of the present application.
Fig. 3 is a front view of the present application.
Description of element numbers:
1. movable support
11. Cavity cavity
12. Baffle plate
13. Sliding chute
14. Magnetic stripe
15. Grip adjusting part
16. Frame body
2. Fixing support
3. Hinge
Detailed Description
In order to make the objects, technical solutions and advantageous effects of the present application clearer, the present application is further described below with reference to examples. It should be understood that the examples are presented by way of illustration only and are not intended to limit the scope of the application. The test methods used in the following examples are conventional, unless otherwise indicated, and other advantages and effects of the present application will be readily apparent to those skilled in the art from the disclosure herein.
The inventor of the application has invented a laboratory protein development auxiliary device through a large amount of experimental research and study, and completed the application on the basis of the laboratory protein development auxiliary device.
The first aspect of the present application provides a protein development auxiliary device, including movable support 1 and fixed bolster 2 that connect, movable support 1 includes framework portion 16 and cavity 11, is equipped with a plurality of baffles 12 on the movable support 1, and baffle 12 is followed framework portion 16 slides, and the bottom of fixed bolster 2 is equipped with magnetic stripe 14. The present application enables accurate development of a plurality of target strip areas by adjusting the position of the shutter 12 without cutting the protein-loaded film.
As shown in fig. 1, in the protein development auxiliary device provided by the application, a movable bracket 1 and a fixed bracket 2 are connected through a hinge 3, and the movable bracket 1 rotates relative to the fixed bracket 2 by taking a rotating shaft of the hinge 3 as an axle center. When the protein film is not required to be shielded, the movable support 1 can be turned upwards, the hinge 3 is at a fixed rotation angle, the movable support 1 can be prevented from appearing in a developed image or can be directly disassembled, and the movable support is directly adsorbed on the outer side of a machine through a magnetic stripe, so that the movable support is convenient to store; when the nonspecific strip of interference needs to be shielded, the movable support is turned over to cover the nonspecific strip on the protein film, the nonspecific strip is shielded by the baffle 12, the hinge 3 is arranged to enable more choices to be made for protein development, and the movable support 1 can appear or disappear according to the development needs.
As shown in fig. 1, in the protein development auxiliary device provided in the present application, a chute 13 is disposed between the frame portion 16 and the cavity 11, or the chute 13 is disposed on the frame portion 16, and the baffle 12 can slide along the chute 13. Specifically, the chute 13 is provided around the cavity 11. In a specific embodiment of the present application, the chute 13 may be, for example, a flat surface, and the baffle 12 slides and is fixed on the chute 13 by friction. In another embodiment of the present application, the sliding groove 13 may be, for example, a concave surface, and the baffle 12 is provided with a protruding sliding block matched with the concave surface of the sliding groove 13, where the protruding sliding block may drive the baffle 12 to slide relative to the sliding groove 13. In one embodiment of the present application, the number of the sliding grooves 13 is 2 to 4, and the even number of the sliding grooves 13 is that the sliding of the at least one baffle 12 can be maintained by the 2 sliding grooves 13. In a preferred embodiment of the present application, the number of runners 13 is 4, which is arranged such that the barrier 12 can block both the transverse strips of the protein film and the longitudinal lanes of the protein film.
In the protein development auxiliary device provided by the application, the baffle 12 is provided with the holding adjusting part 15 for adjusting the position of the baffle 12. In a specific embodiment of the present application, the holding adjustment portion 15 may be, for example, a protrusion or a groove, or may be in other conventionally understood holding shapes. The baffle 12 is made of opaque material, and one skilled in the art can select a proper material according to actual needs. In the specific embodiment of the present application, the protein development aid is located on the console of the chemiluminescent imager, with the baffle 12 parallel to the console and securable within the cavity 11 due to gravity and friction. When the method is actually applied, when the nonspecific strip exists on the protein film, only the target strip area is exposed by controlling the baffle distance, and the color development result of the target strip is accurately obtained.
In the protein development auxiliary device provided by the application, the magnetic stripe 14 is arranged on the fixed support 2 and is used for being adsorbed on an operation table of a development imaging instrument. Specifically, in one embodiment of the present application, the magnetic stripe 14 on the fixing support 2 is located above the operation table of the developing imager, and another magnetic stripe is located below the operation table of the developing imager and is matched with the magnetic stripe 14 of the fixing support 2, and the two magnetic stripes fix the fixing support 2 on the operation table through magnetic attraction between the two magnetic stripes. The device is suitable for most chemiluminescent imagers of different brands, is fixed on the operating platform of the chemiluminescent imagers through magnetic stripe adsorption, does not cause any damage to the operating platform, and is easy to install and detach.
A second aspect of the present application provides a protein development method, using the aforementioned protein development auxiliary device, comprising the steps of:
1) Placing the protein film in the console of the development imager such that the protein film is located in the cavity 11;
2) The movable bracket 1 is overturned relative to the fixed bracket 2, so that the movable bracket does not shade the protein film, and the white light image carrying the protein film is obtained by image acquisition;
3) The movable bracket 1 is put down to cover the protein film, the baffle 12 is slidingly adjusted according to the position of a marker where the molecular weight of the target protein is positioned, so that the target protein is just exposed out of the position, and then chemiluminescent liquid is added to collect a chemiluminescent image, so that a target protein strip chart is obtained;
4) Fitting the white light map carrying the protein film obtained in the step 2) and the target protein strip map obtained in the step 3) to obtain a target image.
In the protein development method provided by the application, step 1) refers to placing the protein film in the operation table of the development imager so that the protein film is located in the cavity 11. The protein membrane comprises target protein, interference protein and protein marker. The method and the device enable the protein film with the interference protein to only expose the target strip area, and accurately obtain the target strip color development result.
In the protein development method provided by the application, the step 2) is to turn over the movable bracket 1 relative to the fixed bracket 2 so as not to shade the protein film, and perform image acquisition to obtain a white light image carrying the protein film. Wherein, after the movable bracket 1 is overturned, the protein development is not affected. The white light map of the protein film includes an overall map of the protein film.
In the protein developing method provided by the application, step 3) refers to putting down the movable bracket 1 to cover the movable bracket on a protein film, sliding and adjusting the baffle 12 according to the position of a marker where the molecular weight of the target protein is located, so that the baffle is just exposed out of the position where the target protein is located, and then adding a chemiluminescent liquid to collect a chemiluminescent image to obtain a target protein strip chart. Among them, ECL chemiluminescent solutions (Thermo Fisher, A38556; elegance, SQ201; tiansheng, 180-5001) and the like are known in the art as conventional chemiluminescent solutions for protein development. The position of the shutter 12 is adjusted by holding the adjusting portion 15. The method can shade the position of the interference protein according to the requirement, obtain a high-quality whole film picture only containing the target protein and the protein marker, and meet the requirements of experimental drawing and article publishing.
In the protein development method provided by the application, the step 4) refers to fitting the white light image carrying the protein film obtained in the step 2) with the target protein strip image obtained in the step 3) to obtain a target image. Wherein the target image comprises all protein markers and target bands.
The specific implementation process of the device is as follows:
when the protein is developed, firstly, the bin gate of the developing imaging instrument is opened, the fixed support 2 of the device is fixed on the operating platform of the developing imaging instrument through the magnetic stripe 14, the protein film is placed on the operating platform, the protein film is positioned in the cavity 11, the movable support 1 is lifted, and the bin gate is closed to collect the white light image of the whole film, so that the white light image carrying the protein film is obtained.
According to the molecular weight of target protein and the position of a marker in a white light image of a protein film, a movable bracket 1 of the device is covered on the protein film, the position of a baffle plate 12 is adjusted by holding an adjusting part 15 to enable the baffle plate to just expose a target strip area, and a natural energy ECL chemiluminescent liquid (180-5001) chemiluminescent liquid is added for image acquisition to obtain the target protein strip image.
Fitting the white light image carrying the protein film and the target protein band image to finally obtain a target band result with an intact protein marker.
In summary, the present application enables accurate development of multiple target strip areas by adjusting the baffle positions without cutting the protein-loaded film. When the used antibody has nonspecific stripes, only the target stripe area is exposed by controlling the baffle distance, and the target stripe color development result is accurately obtained. The device is suitable for most chemiluminescent imagers of different brands, is fixed on the operating platform of the chemiluminescent imagers through magnetic stripe adsorption, does not cause any damage to the operating platform, and is easy to install and detach.
The above embodiments are merely illustrative of the principles of the present invention and its effectiveness, and are not intended to limit the application. Modifications and variations may be made to the above-described embodiments by those skilled in the art without departing from the spirit and scope of the present application. Accordingly, it is intended that all equivalent modifications and variations which can be accomplished by persons skilled in the art without departing from the spirit and technical spirit of the present disclosure shall be covered by the claims of this application.
Claims (10)
1. The utility model provides a protein development auxiliary device, its characterized in that, including movable support (1) and fixed bolster (2) of connection, movable support (1) are including framework portion (16) and cavity (11), be equipped with a plurality of baffles (12) on movable support (1), baffle (12) are followed framework portion (16) slip, the bottom of fixed bolster (2) is equipped with magnetic stripe (14).
2. Protein development aid according to claim 1, characterized in that the movable support (1) and the fixed support (2) are connected by means of a hinge (3).
3. Protein development aid according to claim 1, characterized in that a chute (13) is provided between the frame part (16) and the cavity (11), or that a chute (13) is provided on the frame part, the baffle (12) being slidable along the chute (13).
4. A protein development aid as claimed in claim 3, characterized in that the chute 13 is provided around the cavity (11).
5. Protein development auxiliary device according to claim 1, characterized in that the shutter (12) is provided with a grip adjustment (15).
6. Use of the protein development aid according to any one of claims 1 to 5 for protein development.
7. A protein development method using the protein development auxiliary device according to any one of claims 1 to 5, comprising the steps of:
1) Placing a protein film in a console of a development imager such that the protein film is located in the cavity (11);
2) Turning the movable bracket (1) relative to the fixed bracket (2) to enable the movable bracket not to shade the protein film, and collecting images to obtain a white light image carrying the protein film;
3) Putting down the movable bracket (1) to cover the protein film, sliding and adjusting the baffle (12) according to the position of a marker where the molecular weight of the target protein is located, so that the baffle just exposes the position where the target protein is located, and then adding chemiluminescent liquid to collect a chemiluminescent image to obtain a target protein strip chart;
4) Fitting the white light map carrying the protein film obtained in the step 2) and the target protein strip map obtained in the step 3) to obtain a target image.
8. The method of developing proteins of claim 7, wherein in step 1), the protein film comprises a protein of interest, an interfering protein, and a protein marker.
9. The protein developing method according to claim 7, wherein in step 3), the position of the shutter (12) is adjusted by holding an adjusting portion (15).
10. The protein development method of claim 7, wherein in step 4), the target image comprises a protein marker and a target protein.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310436906.7A CN116500275A (en) | 2023-04-21 | 2023-04-21 | Laboratory protein development auxiliary device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310436906.7A CN116500275A (en) | 2023-04-21 | 2023-04-21 | Laboratory protein development auxiliary device |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116500275A true CN116500275A (en) | 2023-07-28 |
Family
ID=87322396
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310436906.7A Pending CN116500275A (en) | 2023-04-21 | 2023-04-21 | Laboratory protein development auxiliary device |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116500275A (en) |
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2023
- 2023-04-21 CN CN202310436906.7A patent/CN116500275A/en active Pending
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