CN107192815B - A kind of antibody incubation kit and its application method - Google Patents
A kind of antibody incubation kit and its application method Download PDFInfo
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- CN107192815B CN107192815B CN201710442993.1A CN201710442993A CN107192815B CN 107192815 B CN107192815 B CN 107192815B CN 201710442993 A CN201710442993 A CN 201710442993A CN 107192815 B CN107192815 B CN 107192815B
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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
- G01N33/531—Production of immunochemical test materials
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Abstract
The invention discloses a kind of antibody incubation kit and its application method, the antibody incubation kit includes box body and box cover, and the box body includes silicon rubber, waterproof film, wet tissue and groove；Silicon rubber is housed, waterproof film is laid on silicon rubber, is arranged fluted on waterproof film, and the inner wall of box body is with wet tissue in the box body.Application method are as follows: the groove lower than silicon rubber plane is produced on the waterproof film for being laid in silicon rubber, after hybond membrane is carried out transferring film, rinsing, hybond membrane protein-contg one faces upward, it is horizontally arranged in groove, antibody diluent is added dropwise, it is infiltrated on hybond membrane in antibody diluent, carries out hybridization reaction.The application method of antibody incubation kit of the invention is easy to operate, effect is good, at low cost.
Description
Technical field
The invention belongs to experimental apparatus fields, and in particular to a kind of antibody incubation kit and its application method.
Background technique
Commonly using the inspection of the hybridization blot assays such as western blotting, southern blotting in Bioexperiment
Survey purpose band.The incubation process of hybridization, conventional method are that the film where molecule is fully immersed in liquid antibody-containing,
Time by > 1 hour makes antibody in conjunction with the molecules of interest on film.
In Western blotting, generally film and antibody are placed in capsule, are incubated on shaking table, the method is required every time
Amount of antibody it is larger；It is difficult to cope with multispecific antibody incubation；Antibody recycling is not exclusively, different according to the material of capsule, the efficiency of recycling
It is not quite similar.The method of another kind antibody incubation at present, is to be placed in film and antibody in the pouch that plastic packaging film is made into, the method exists
A large amount of wastes of time and antibody are caused when sample-adding and antibody recycling, and are difficult to remove the bubble for having extreme influence to experiment；
In addition plastic packaging film is disposably used, is resulted in waste of resources and environmental pollution.
Therefore it is necessary to provide kit and its user of the convenient and efficient, effect stability of one kind and save the cost
Method.
Summary of the invention
The purpose of the present invention is to provide a kind of antibody incubation kit easy to operate, effect is good, at low cost and its make
Use method.
In order to achieve the object of the present invention, the present invention provides a kind of antibody incubation kits, described comprising box body and box cover
Box body includes silicon rubber, waterproof film, wet tissue and groove；Silicon rubber is housed, waterproof film is laid in silicon in the box body
On rubber, it is arranged fluted on waterproof film, the inner wall of box body is with wet tissue.
Further, the silicon rubber is Polymethyl methacrylate.
Further, thickness >=5mm of the silicon rubber.
Further, the waterproof film is masking foil, preservative film or sealed membrane.
Further, the quantity of the groove is more than one.
Further, the groove is polygon, round or irregular shape；Preferably, the groove is to agree with hybond membrane
Polygon, round or irregular shape.
The present invention also provides a kind of application methods of above-mentioned antibody incubation kit: being laid in the waterproof of silicon rubber
Produce the groove lower than silicon rubber plane on film, hybond membrane be subjected to transferring film, after rinsing, hybond membrane protein-contg one towards
On, it is another to be horizontally arranged in groove towards bottom portion of groove, antibody diluent is added dropwise, is infiltrated on hybond membrane in antibody diluent,
Carry out hybridization reaction.
Further, the groove is formed by applying certain pressure on waterproof film, makes waterproof film and groove
It fits closely, the depth of groove is 2-3mm, and groove area is greater than the area of hybond membrane.
Further, the additive amount of the antibody diluent is 3~4mL antibody diluent/(6cm × 8cm) hybond membrane.
Further, the hybond membrane is pvdf membrane or NC film.
Under normal circumstances, experiment, which is incubated for 6cm × 8cm hybond membrane using capsule, needs 10ml or more antibody diluent, the present invention
The additive amount of antibody diluent is 3~4mL antibody diluent/(6cm × 8cm) hybond membrane, greatly reduces antibody dosage, effectively
Reagent cost is saved.
Antibody incubation kit of the present invention is with a circle wet tissue in inboard wall of cartridge, and the steaming of dilution can be effectively prevented
Hair.
The present invention utilizes the mobility and plasticity of silicon rubber, and the waterproof thin film fabrication for making to be laid on silicon rubber is low out
In the groove of silicon rubber plane, for providing space to antibody incubation.
Kit simple and effective of the present invention, application method is simple, and multiple grooves can be set as needed, and carries out simultaneously
The incubation of different antibodies.Groove can make according to the size and shape of hybond membrane, can be round, polygon or irregular figure,
It is formed by that space is minimum, the application of just right probe dilution liquid is greatly saved amount of antibody.
When being incubated for Multiple Antibodies, multiple grooves can be made in the silicon rubber plane in the same seal box, while into
Row experiment.Antibody is only stored in groove, no cross contamination.Shaking table can be placed to be incubated for obtain better crossbreeding effect.
Due to foring certain groove, when being incubated on shaking table, antibody diluent be will not flow out, and it is good to be incubated for effect.
Groove is convenient for the direct recycling of antibody, the in addition hydrophobicity of waterproof film, therefore antibody kit of the invention
Antibody diluent the rate of recovery it is good, up to 99%.
The application method of antibody incubation kit of the present invention with hermetic bag be incubated for antibody method compared with, take pvdf membrane and
It is more convenient and complete to recycle antibody, has saved twice or more of time cost and reagent cost.After use all utensils and
Material rear recycling capable of washing.
It is miscellaneous that antibody incubation kit of the present invention can be used for the traces such as western blotting, southern blotting
Hand over experiment.
Detailed description of the invention
Fig. 1 is kit top view；
Wherein: box body 1, silicon rubber 2, wet tissue 3, waterproof film 4, groove 5, hybond membrane 6.
Specific embodiment
The embodiment of the present invention is described below in detail, the examples of the embodiments are intended to be used to explain the present invention, and cannot
It is interpreted as limitation of the present invention.In the examples where no specific technique or condition is specified, described according to the literature in the art
Technology or conditions or carried out according to product description.Reagents or instruments used without specified manufacturer is that can lead to
Cross the conventional products of commercially available acquisition.
The preparation of 1 antibody kit of embodiment
1, a sealable box is taken, the silicon rubber 2 for being laid with 5mm thickness inside it makes to form a water in box body 1
Plane.
2, the waterproof film 4 that length and width is slightly larger than pvdf membrane 6 is cut, it is flat that waterproof film 4 is arranged in silicon rubber 2
Suitable position tiling on face.Using the plasticity of Polymethyl methacrylate, applies to waterproof 4 Middle face of film and press
Power, so that bottom surface sinking 2-3mm.So far waterproof 4 bottom of film forms groove 5 at plane, and groove 5 and waterproof film 4 are tight
Closely connected conjunction.
3, it makes a circle wet tissue 3 in seal box inner wall, to prevent the evaporation of antibody diluent.
Waterproof film can be sealed membrane, preservative film or masking foil in embodiment.
Silicon rubber is Polymethyl methacrylate in embodiment.
The use of 2 antibody kit of embodiment
By taking western blotting as an example:
6 transferring film of hybond membrane, it is rinsed after, pvdf membrane is faced upward containing albumen one, is horizontally arranged in 1 kit of embodiment
Groove 5 in, directly dropwise addition antibody diluent.The additive amount of antibody diluent is 3~4mL antibody diluent/(6cm × 8cm)
Hybond membrane.The volume of antibody diluent should not exceed groove 5.It is infiltrated on hybond membrane 6 in antibody diluent, hybridize anti-
It answers.Incubation process can keep horizontal positioned, can also be slowly shaken with low-angle.
Antibody recycling: lift one jiao take hybond membrane away after, antibody-solutions reside in the groove 5 that waterproof film 4 is formed,
It directly can quickly be drawn using liquid-transfering gun.
Hybond membrane is pvdf membrane or NC film in embodiment.
The experiment proved that the antibody diluent of the method returns without causing antibody to be lost because of misoperation
Yield is up to 99%.It is recycled after utensil and material used in the method are capable of washing, saves consumables cost.
Although the embodiments of the present invention has been shown and described above, it is to be understood that above-described embodiment is example
Property, it is not considered as limiting the invention, those skilled in the art are not departing from the principle of the present invention and objective
In the case where can make changes, modifications, alterations, and variations to the above described embodiments within the scope of the invention.
Claims (10)
- It include box body and box cover 1. a kind of antibody incubation kit, which is characterized in that the box body includes silicon rubber (2), impermeable Water film (4), wet tissue (3) and groove (5)；Silicon rubber (2) are housed in the box body (1), waterproof film (4) is laid in silicon rubber On glue (2), it is arranged fluted (5) on waterproof film (4), the inner wall of box body (1) is with wet tissue (3)；Groove (5) basis The size and shape of hybond membrane makes, and by applying pressure initiation on waterproof film (4), makes waterproof film (4) bottom At plane, waterproof film (4) fits closely with groove；Antibody diluent in the groove are as follows: every 6cm × 8cm hybond membrane adds Add 3 ~ 4 mL antibody diluents.
- 2. antibody incubation kit according to claim 1, which is characterized in that the silicon rubber (2) is poly- methyl ethylene Siloxanes.
- 3. antibody incubation kit according to claim 1, which is characterized in that thickness >=5mm of the silicon rubber (2).
- 4. antibody incubation kit according to claim 1, which is characterized in that the waterproof film (4) is masking foil, protects Fresh film or sealed membrane.
- 5. antibody incubation kit according to claim 1, which is characterized in that the quantity of the groove (5) is more than one.
- 6. antibody incubation kit according to claim 1, which is characterized in that the groove (5) is polygon, round or not Regular shape.
- 7. antibody incubation kit according to claim 6, which is characterized in that the groove (5) is to agree with hybond membrane (6) Polygon, round or irregular shape.
- 8. a kind of application method of any one of claim 1-7 antibody incubation kit, which is characterized in that be laid in silicon The groove (5) lower than silicon rubber (2) plane is produced on the waterproof film (4) of rubber (2), by hybond membrane (6) carry out transferring film, After rinsing, hybond membrane (6) protein-contg one faces upward, another to be horizontally arranged in groove (5) towards groove (5) bottom, is added dropwise anti- Body dilution is infiltrated on hybond membrane (6) in antibody diluent, carries out hybridization reaction.
- 9. the application method of antibody incubation kit according to claim 8, which is characterized in that the depth of the groove (5) For 2-3mm, groove (5) area is greater than the area of hybond membrane (6).
- 10. according to the application method of any one of the claim 8-9 antibody incubation kit, which is characterized in that the hybridization Film is pvdf membrane or NC film.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710442993.1A CN107192815B (en) | 2017-06-13 | 2017-06-13 | A kind of antibody incubation kit and its application method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710442993.1A CN107192815B (en) | 2017-06-13 | 2017-06-13 | A kind of antibody incubation kit and its application method |
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| Publication Number | Publication Date |
|---|---|
| CN107192815A CN107192815A (en) | 2017-09-22 |
| CN107192815B true CN107192815B (en) | 2019-06-11 |
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| CN201710442993.1A Active CN107192815B (en) | 2017-06-13 | 2017-06-13 | A kind of antibody incubation kit and its application method |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN204422535U (en) * | 2015-01-13 | 2015-06-24 | 河南赛诺特生物技术有限公司 | A kind of novel agent box possessing the immunohistochemical reagents of moisture-keeping functions |
| CN205620419U (en) * | 2016-02-16 | 2016-10-05 | 张新化 | Western blotting determine module |
| CN106093372A (en) * | 2016-06-07 | 2016-11-09 | 遵义医学院 | A kind of western blot antibody incubation box |
| CN206103954U (en) * | 2016-10-14 | 2017-04-19 | 湖州数康生物科技有限公司 | Biochip box that high flux jointly detected or filtered based on silica gel matrix |
| CN106610429A (en) * | 2015-10-26 | 2017-05-03 | 宁波大学 | Dual-drive tumor marker joint inspection chip device with PDMS as substrate |
-
2017
- 2017-06-13 CN CN201710442993.1A patent/CN107192815B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN204422535U (en) * | 2015-01-13 | 2015-06-24 | 河南赛诺特生物技术有限公司 | A kind of novel agent box possessing the immunohistochemical reagents of moisture-keeping functions |
| CN106610429A (en) * | 2015-10-26 | 2017-05-03 | 宁波大学 | Dual-drive tumor marker joint inspection chip device with PDMS as substrate |
| CN205620419U (en) * | 2016-02-16 | 2016-10-05 | 张新化 | Western blotting determine module |
| CN106093372A (en) * | 2016-06-07 | 2016-11-09 | 遵义医学院 | A kind of western blot antibody incubation box |
| CN206103954U (en) * | 2016-10-14 | 2017-04-19 | 湖州数康生物科技有限公司 | Biochip box that high flux jointly detected or filtered based on silica gel matrix |
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| Publication number | Publication date |
|---|---|
| CN107192815A (en) | 2017-09-22 |
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