CN201662574U - Lung cancer immunoblotting reagent box - Google Patents
Lung cancer immunoblotting reagent box Download PDFInfo
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- CN201662574U CN201662574U CN2010201195095U CN201020119509U CN201662574U CN 201662574 U CN201662574 U CN 201662574U CN 2010201195095 U CN2010201195095 U CN 2010201195095U CN 201020119509 U CN201020119509 U CN 201020119509U CN 201662574 U CN201662574 U CN 201662574U
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- lung cancer
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- immunoblotting
- reagent box
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Abstract
The utility model relates to a lung cancer immunoblotting reagent box, which is characterized by comprising a box body, a lung cancer tissue blotting protein membrane (2), membrane storage liquor (3) and concentrated membrane cleaning liquor (4). The membrane storage liquor (3) and the concentrated membrane cleaning liquor (4) are respectively in bottled structures and fixed into the box body (1) via paper templates, and the lung cancer blotting tissue protein membrane (2) is packed in a storage bag (5) in a sealing manner. The lung cancer tissue blotting protein membrane of the reagent box as a technical core is formed by the aid of lung cancer tissue protein extracting, electrophoretic separation and membrane conversion preparation, is extremely convenient and speedy in both storage and transportation, and can be used for a plurality of times, thereby the lung cancer immunoblotting reagent box realizes highly efficient combination and utilization of lung cancer sample resources and research.
Description
Technical field
The utility model relates to a kind of lung cancer HP immunoblotting kit, belongs to the medical research field.
Background technology
Lung cancer claims lung bronchogenic carcinoma again, is the malignant tumour that originates from bronchus and alveolar epithelial cells, lung cancer become present M ﹠ M increase the fastest, human health and life are threatened maximum malignant tumour.5 years survival rates of lung cancer only have 10%～15%, and the treatment of advanced lung cancer is difficult point especially.80% is non-small cell lung cancer in the lung cancer, and based on squama cancer, gland cancer and large cell carcinoma, has been middle and advanced stage during most the discovery, and combined therapy effects such as operation, chemicotherapy are undesirable.In addition, 20% lung cancer is small-cell carcinoma of the lung, and its overall prognosis is poorer.Therefore, very urgent for the research of lung cancer.Along with developing rapidly of Protocols in Molecular Biology, the research of lung cancer has been deep into gene, albumen equimolecular level, its pathogenesis also is familiar with gradually and is pushed to utilization.Western blotting (immunoblotting) claims Western blotting (western blotting) again, and it is the method according to certain albumen in the specificity binding characteristic detection of complex sample of antigen-antibody.High specific and susceptibility that it has high resolution and the solid-phase immunoassay of SDS-PAGE are usually used in identifying certain albumen, and can carry out qualitative and semi-quantitative analysis to albumen.The Western blotting technology is widely used in physianthropy research.Lung cancer is as tumour occurred frequently, is indispensable ingredient in the present lung cancer research field to the research of its associated protein.
The utility model content
It is a kind of convenient and swift, workable that the purpose of this utility model is to provide, and the lung cancer HP immunoblotting kit of good reproducibility.
The purpose of this utility model can reach by the following technical programs:
A kind of lung cancer HP immunoblotting kit, comprise box body, cancerous lung tissue trace albumen diaphragm, film storage liquid and concentrated film washing liquid, described film storage liquid and concentrated film washing liquid are respectively bottled structure, be fixed in the box body by the papery template, the diaphragm seal of described lung cancer trace histone is loaded in the storage bag.
The purpose of this utility model can also reach by the following technical programs:
A kind of embodiment of the present utility model is: described film storage liquid can be 0.02% Sodium azide-TBST solution, totally 1 bottle.
A kind of embodiment of the present utility model is: described concentrated film washing liquid can be damping fluid, totally 1 bottle, and capacity 1L.
A kind of embodiment of the present utility model is: described storage bag can be the plastics hybridization bag.
The utlity model has following outstanding beneficial effect:
Cancerous lung tissue trace albumen diaphragm of the present utility model is a technological core, it is the extracting, electrophoretic separation by lung cancer tissue protein and changes film preparation and form, preserve with transportation all very convenient quick, and can repeatedly use, thereby the high effective integration and the utilization of lung cancer specimen resource and research have been realized.
Description of drawings
Fig. 1 is a structural representation of the present utility model.
Wherein, the 1-box body, 2-cancerous lung tissue trace albumen diaphragm, 3-stores liquid, and 4-concentrates film washing liquid, the 5-storage bag.
Embodiment
Embodiment 1:
With reference to Fig. 1, present embodiment comprises box body 1, cancerous lung tissue trace albumen diaphragm 2, film storage liquid 3 and concentrated film washing liquid 4, film storage liquid 3 and concentrated film washing liquid 4 are bottled structure, are fixed in the box body 1 by the papery template, and described lung cancer trace histone diaphragm 2 sealings are loaded in the storage bag 5.
In the present embodiment,
The preparation process of cancerous lung tissue trace albumen diaphragm is as follows:
(1) extracting of albumen
At first, cut an amount of cancerous lung tissue, remove bloodstain and charcoal dirt with the damping fluid rinsing, blot and shred and put into homogenizer, add radioimmunoprecipitation assay cracking liquid and protease inhibitors in homogenizer, then, freeze thawing homogenate tissue is 3 times rapidly, make the abundant cracking of homogenate product, pyrolysis product is transferred in the centrifuge tube fully, under 4 ℃,, gets supernatant and be sub-packed in the centrifuge tube with centrifugal 50min, part places-80 ℃ of preservations, and part is used for determination of protein concentration;
(2) protein sample pre-service
The extracting protein sample mixes by 1: 1 volume ratio with the 2X sample-loading buffer, after boiling water boils, puts ice bath immediately, and temperature is reduced to about room temperature, mild centrifugation then, and vortex is even, and is centrifugal again, sample in the preparation;
(3) electrophoretic separation
At first prepare separation gel and concentrated glue, the wherein preparation of separation gel, composition is: ultrapure water, the two third rare acid amides of rare acid amides/methene, Tri(Hydroxymethyl) Amino Methane Hydrochloride, lauryl sodium sulfate, Ammonium Persulfate 98.5 and tetramethyl diethylamine, and the preparation composition that concentrates glue is: ultrapure water, the two third rare acid amides of third rare acid amides/methene, Tri(Hydroxymethyl) Amino Methane Hydrochloride, lauryl sodium sulfate, Ammonium Persulfate 98.5 and tetramethyl diethylamine; Separation gel and concentrated glue are placed in the mould, add fresh electrophoretic buffer, survey not have leak back polishing electrophoretic buffer, then on the gel both sides sample hole add and pre-dsred protein molecular weight standard product, electrophoresis runs at the bottom of the glue until the bromophenol blue dyestuff;
(5) change film
Open glass plate, strike off concentrated glue, take out separation gel, discharge is changeed liquid and is soaked, PVDF membrane and filter paper have been sheared by the glue size again, make sandwich then, the positive glue of film is negative, and promptly negative pole-clip-foam-rubber cushion-filter paper-gel-PVDF membrane-filter paper-foam-rubber cushion-clip-positive pole is put into transfer groove with sandwich, make the black flour of the black flour of folder to groove, the fine flour of folder is to red of groove, and the polishing electricity changes liquid, carries out 2h with constant voltage 100V, protein transduction is moved on the PVDF membrane, can make lung cancer tissue protein blotting membrane.
The preparation method of kit is as follows:
As shown in Figure 1, described lung cancer trace histone diaphragm 2 sealings are loaded in the storage bag 5, add an amount of film storage liquid 3, store down at 4 ℃.
The prescription of film storage liquid: 0.02% Sodium azide is dissolved in TBST (trishydroxymethylaminomethane 3g, hydrochloric acid 1.65ml, Tween-20 1ml is settled to 1000ml, regulating PH is 7.5 for sodium chloride 8g, potassium chloride 0.2g)
Concentrate the prescription of film washing liquid (10X): 12.11g trishydroxymethylaminomethane (Tris), 87.66g sodium chloride (NaCl), 5ml non-ionic surfactant (Tween-20), 1g Sodium azide, PH are 7.4.Before use with 10 times of deionized water dilutions.
Claims (4)
1. lung cancer HP immunoblotting kit, it is characterized in that: comprise box body (1), cancerous lung tissue trace albumen diaphragm (2), film storage liquid (3) and concentrated film washing liquid (4), described film storage liquid (3) and concentrated film washing liquid (4) are respectively bottled structure, be fixed in the box body (1) by the papery template, described lung cancer trace histone diaphragm (2) sealing is loaded in the storage bag (5).
2. lung cancer HP immunoblotting kit according to claim 1 is characterized in that: described film storage liquid (3) is 0.02% Sodium azide-TBST solution.
3. lung cancer HP immunoblotting kit according to claim 1 is characterized in that: described concentrated film washing liquid (4) is a damping fluid.
4. lung cancer HP immunoblotting kit according to claim 1 is characterized in that: described storage bag (5) is a polybag.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010201195095U CN201662574U (en) | 2010-02-10 | 2010-02-10 | Lung cancer immunoblotting reagent box |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010201195095U CN201662574U (en) | 2010-02-10 | 2010-02-10 | Lung cancer immunoblotting reagent box |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN201662574U true CN201662574U (en) | 2010-12-01 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010201195095U Expired - Fee Related CN201662574U (en) | 2010-02-10 | 2010-02-10 | Lung cancer immunoblotting reagent box |
Country Status (1)
| Country | Link |
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| CN (1) | CN201662574U (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112730847A (en) * | 2020-12-22 | 2021-04-30 | 哈尔滨赛信生物科技开发有限公司 | Kit for immunoblotting experiment by using antibody and application thereof |
-
2010
- 2010-02-10 CN CN2010201195095U patent/CN201662574U/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112730847A (en) * | 2020-12-22 | 2021-04-30 | 哈尔滨赛信生物科技开发有限公司 | Kit for immunoblotting experiment by using antibody and application thereof |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20101201 Termination date: 20120210 |