CN109425742A - A kind of chemoluminescence method quickly detects the kit of aquaporin 1 - Google Patents
A kind of chemoluminescence method quickly detects the kit of aquaporin 1 Download PDFInfo
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- CN109425742A CN109425742A CN201710732285.1A CN201710732285A CN109425742A CN 109425742 A CN109425742 A CN 109425742A CN 201710732285 A CN201710732285 A CN 201710732285A CN 109425742 A CN109425742 A CN 109425742A
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- 102000004888 Aquaporin 1 Human genes 0.000 title abstract description 19
- 108090001004 Aquaporin 1 Proteins 0.000 title abstract description 19
- 238000000034 method Methods 0.000 title abstract description 13
- 102000010637 Aquaporins Human genes 0.000 abstract description 22
- 108010063290 Aquaporins Proteins 0.000 abstract description 22
- 239000007788 liquid Substances 0.000 abstract description 16
- 108090000790 Enzymes Proteins 0.000 abstract description 14
- 102000004190 Enzymes Human genes 0.000 abstract description 14
- 239000007790 solid phase Substances 0.000 abstract description 11
- 239000000758 substrate Substances 0.000 abstract description 11
- 210000002700 urine Anatomy 0.000 abstract description 6
- 238000003745 diagnosis Methods 0.000 abstract description 4
- 239000002075 main ingredient Substances 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 20
- 229940088598 enzyme Drugs 0.000 description 11
- 238000001514 detection method Methods 0.000 description 8
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 7
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 6
- 235000005979 Citrus limon Nutrition 0.000 description 6
- 244000131522 Citrus pyriformis Species 0.000 description 6
- 210000002966 serum Anatomy 0.000 description 6
- 206010028980 Neoplasm Diseases 0.000 description 5
- 102100037896 Perilipin-2 Human genes 0.000 description 5
- 239000000872 buffer Substances 0.000 description 5
- 108010067163 Perilipin-2 Proteins 0.000 description 4
- 208000006265 Renal cell carcinoma Diseases 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 210000003734 kidney Anatomy 0.000 description 4
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- AQLJVWUFPCUVLO-UHFFFAOYSA-N urea hydrogen peroxide Chemical compound OO.NC(N)=O AQLJVWUFPCUVLO-UHFFFAOYSA-N 0.000 description 4
- 108090001008 Avidin Proteins 0.000 description 3
- LDKDGDIWEUUXSH-UHFFFAOYSA-N Thymophthalein Chemical compound C1=C(O)C(C(C)C)=CC(C2(C3=CC=CC=C3C(=O)O2)C=2C(=CC(O)=C(C(C)C)C=2)C)=C1C LDKDGDIWEUUXSH-UHFFFAOYSA-N 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 229960002685 biotin Drugs 0.000 description 3
- 235000020958 biotin Nutrition 0.000 description 3
- 239000011616 biotin Substances 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 230000002596 correlated effect Effects 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 238000002372 labelling Methods 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
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- 208000009060 clear cell adenocarcinoma Diseases 0.000 description 2
- 210000004408 hybridoma Anatomy 0.000 description 2
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- 239000004005 microsphere Substances 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 210000004988 splenocyte Anatomy 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 206010003445 Ascites Diseases 0.000 description 1
- 108091006146 Channels Proteins 0.000 description 1
- 102000034573 Channels Human genes 0.000 description 1
- 208000030808 Clear cell renal carcinoma Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 206010061183 Genitourinary tract neoplasm Diseases 0.000 description 1
- 101000684063 Homo sapiens Aquaporin-1 Proteins 0.000 description 1
- 101001096050 Homo sapiens Perilipin-2 Proteins 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 1
- 208000008385 Urogenital Neoplasms Diseases 0.000 description 1
- 239000002696 acid base indicator Substances 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000002038 chemiluminescence detection Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 102000052557 human AQP1 Human genes 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 229940127121 immunoconjugate Drugs 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 201000010982 kidney cancer Diseases 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
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- 238000012986 modification Methods 0.000 description 1
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- 239000000178 monomer Substances 0.000 description 1
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- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 201000010279 papillary renal cell carcinoma Diseases 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
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- 238000002601 radiography Methods 0.000 description 1
- 230000036632 reaction speed Effects 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
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- 230000000630 rising effect Effects 0.000 description 1
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- 238000000926 separation method Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 210000005239 tubule Anatomy 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 230000002485 urinary effect Effects 0.000 description 1
Classifications
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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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Hematology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
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Abstract
The object of the invention is used as auxiliary diagnosis to use in providing a kind of kit for quickly detecting aquaporin 1 based on chemoluminescence method, for detecting aquaporin 1 in human urine sample.A kind of 1 chemoluminescence method kit of people's aquaporin, including sample treatment solution, enzyme standard liquid, the solid phase carrier for being coated with capture antibody, luminous substrate it is characterized by: the capture antibody is anti-human 1 polyclonal antibody of aquaporin；The enzyme standard liquid, main ingredient are that enzyme marks anti-human 1 monoclonal antibody of aquaporin.
Description
Technical field
The invention belongs to biological medicine technology chemoluminescence method detection fields, relate in particular to a kind of based on chemiluminescence
Detection people's aquaporin 1 detection kit.
Background technique
Clear-cell carcinoma (abbreviation kidney) is one of the higher tumour of grade of malignancy and the most common tumour in urinary system,
It is initiated by the malignant tumour of kidney essence uriniferous tubule epithelial systems, also known as Grawitz's tumor, accounts for the 80%~90% of kidney malignant tumour.
According to investigations, kidney accounts for second in China's urogenital neoplasm, is only second to tumor of bladder, accounts for adult malignancies'
2%~3%, 20% or so of Children Malignant Tumor, in recent years disease incidence still stable rising.
Current study show that 5 years survival rates 85% of discovery treatment, middle and advanced stage lump are larger when kidney initial stage lump is smaller
Shi Faxian treats 5 years survival rates and drops to 62%, and 5 years survival rates are further reduced to 20% after nodule transfer.It can be seen that early stage
It was found that it is significant for improving survival, it is particularly important for improving prognosis of patients with renal cell carcinoma.
So far, Diagnosis of Renal Cell Carcinoma early stage generally uses B ultrasound, and CT, X-ray radiography etc. can find to diagnose for 1cm or more, right
It is difficult to differentiate between in too small lump, it is existing research shows that aquaporin 1 (Aquaporin, AQP1) and perilipin 2 in urine
(perilipin-2, PLIN2, ADFP) concentration and transparent clear-cell carcinoma, Papillary Renal Cell Carcinoma patient are higher related, concentration with
Tumor size is related to the stage, there is great meaning for diagnosis and treatment, can be fast in clinic there is an urgent need in the art to provide one kind
The method that speed accurately detects aquaporin 1 and perilipin 2, to meet clinical needs.
Summary of the invention
The object of the invention is in providing a kind of kit for quickly detecting aquaporin 1 based on chemoluminescence method, for examining
Aquaporin 1 in human urine sample is surveyed to use as auxiliary diagnosis.
In order to achieve the above object, present invention employs following technical steps:
A kind of 1 chemoluminescence method kit of people's aquaporin, including sample treatment solution, containing enzyme mark anti-human aquaporin
1 monoclonal antibody enzyme standard liquid of albumen, the solid phase carrier for being coated with anti-human 1 polyclonal antibody of aquaporin, luminous substrate.
1 monoclonal antibody of anti-human aquaporin can be exempted from by user's aquaporin 1 as immunogene
Epidemic disease animal is made, preferably source of mouse or rabbit source.1 polyclonal antibody of anti-human aquaporin, can be logical by user's water
Road albumen 1 is made as immunogen immune animal, preferably source of mouse, Ma Yuan, Yang Yuan, rabbit or cavy source.It is above-mentioned to be used as immunogene
The recombined human aquaporin 1 that can be obtained by gene work eukaryotic expression cloned gene of people's aquaporin 1, can also be with
People's aquaporin 1 is obtained using culture people's renal proximal tubular cell purifying.It cultivates the purifying of people's renal proximal tubular cell and obtains people
Aquaporin 1 can be the complete tetramer of people's aquaporin 1, can also be one of any by four monomers being hydrolyzed into
Or combination.
Enzyme mark in the enzyme standard liquid directly can mark aforementioned anti-human aquaporin using horseradish peroxidase mutase
Avidin label horseradish peroxidase mutase and the anti-human aquaporin of biotin labeling also can be used in 1 monoclonal antibody of albumen
1 monoclonal antibody monoclonal antibody of albumen combines the conjugate formed, anti-human using horseradish peroxidase mutase-Avidin-Biotin-
The concentration that enzyme can be improved when 1 monoclonal antibody conjugates of aquaporin expands the amplification factor of entire luminescence-producing reaction, reaction
Intensity accelerates reaction speed.
The sample treatment solution, can be used PBS, PBST, CB, Tris, EDTA, and the those skilled in the art such as TE commonly use
The buffer of pH6~10, directly directly can also use sample without sample treatment solution, directly use can exist when sample it is higher
Detection background detection background values can be effectively reduced using the buffer of pH6.0~10.0, use the CB buffer of pH9.6
When, detection signal-to-noise ratio is apparently higher than the buffer of pH6.0~8.0, the preferred pH9.4~pH10.0 of sample treatment solution.For more preferable family
True buffer is located at most suitable pH value and acid-base indicator, preferred thymolphthalein in the present invention is added.
The solid phase carrier, which can be used the art such as ELISA Plate, polystyrene microsphere, magnetic microsphere and commonly use, exempts from
Epidemic disease adsorption solid phase material.
The luminous substrate can using commercialization luminous substrate, also can be used luminol or its by modification after
Peroxide agent of the compound as the horseradish peroxidases such as reducing agent and hydrogen peroxide, urea peroxide mutase sensitivity, reducing agent
It can be dispensed in same component or independently with oxidant, luminous substrate long-time stability can be improved in independent packing.
Specific embodiment
Embodiment 1: 1 Antibody preparation of water resistant channel protein
People's renal proximal tubular cell is cultivated, purifying obtains 4 segment fraction of polymer of natural aquaporin 1 after cracking, makees
For immunogene 1；Natural 1 list of aquaporin is obtained using separation after pancreatin hydrolysis to 4 segment fraction of polymer of natural aquaporin 1
Body is as immunogene 2；The people's aquaporin 1 obtained by genetic recombination eukaryotic expression cloned gene, as immunogene 3,
Add Freund's complete adjuvant that mouse is immunized using three kinds of immunogenes respectively, it is immune by enhancing three times, it is anti-to extract serum measurement target
Body potency purifies serum collected by AGP test potency up to the whole serum of acquisition after execution mouse after 1: 16
Polyclonal antibody is obtained, while spleen being taken to prepare splenocyte suspension, is added after taking murine myeloma cell to mix with splenocyte suspension
Enter polyethylene glycol, cultivated after fusion through selectivity and obtain hybridoma, screens secreting specificity antibody hybridoma through increasing
It is inoculated in mouse peritoneal after growing, obtains monoclonal antibody by acquisition mouse ascites purifying in 2 weeks or so.All acquisitions it is polyclonal
Antibody and monoclonal antibody are all positive with former immunogene through diffusion method.Polyclonal antibody potency is better than monoclonal antibody,
The polyclonal antibody and antibody titer that 1 source of immunogene generates are better than polyclonal antibody and Dan Ke that immunogene 2 generates
Grand antibody.It is verified by protein immunoblotting, what the miscellaneous band of polyclonal antibody in 1 source of immunogene was generated more than immunogene 2
Polyclonal antibody.
Embodiment 2:
A kind of 1 chemoluminescence method kit of people's aquaporin, including sample treatment solution, enzyme standard liquid, to be coated with capture anti-
Solid phase carrier, luminous substrate, the standard items of body.
Sample treatment solution ingredient: the CB of pH9.4~10.0 contains 0.1% thymolphthalein.
Enzyme standard liquid main component: 1 antibody of horseradish peroxidase mutase rabbit-anti recombined human aquaporin.
Be coated with the solid phase carrier of capture antibody: the magnetism of coating anti-1 polyclonal antibody of recombined human aquaporin of mouse is micro-
Ball.
Luminous substrate: A liquid is that luminol content is 0.02M, and the TE solution of pH=8.0, B liquid is that every 100mL solution contains lemon
The TE solution of the pH=8.0 of lemon acid 0.282g, 0.2g urea peroxide.
By detecting to 10 parts of patients and 100 parts of Healthy People volunteer's urines, mean value is detected plus 3 marks with volunteer
Quasi- poor, patient's positive rate 80%, Healthy Human Serum negative rate 100%.
1: 10,1: 100,1: 1000,1: 10000,1: 100000,5 are carried out to the positive sample of 1 part of 200ng/ml or more
Gradient is diluted, and is positively correlated through detection luminous intensity with concentration, Fitted logistic curves, R > 0.9.
Embodiment 3:
A kind of 1 chemoluminescence method kit of people's aquaporin, including sample treatment solution, enzyme standard liquid, to be coated with capture anti-
The solid phase carrier of body, luminous substrate.
Sample treatment solution ingredient: the PBS of pH 7.4.
Enzyme standard liquid main component: Avidin marks horseradish peroxidase mutase and the anti-human aquaporin of biotin labeling
1 monoclonal antibody monoclonal antibody combines the conjugate formed.
It is coated with the solid phase carrier of capture antibody: the ELISA Plate of coating anti-1 antibody of recombined human aquaporin of mouse.
Luminous substrate: A liquid is that luminol content is 0.02M, and the TE solution of pH=8.0, B liquid is that every 100mL solution contains lemon
The TE solution of the pH=8.0 of lemon acid 0.282g, 0.2g urea peroxide.
By detecting to 10 parts of patients and 100 parts of Healthy People volunteer's urines, mean value is detected plus 3 marks with volunteer
Quasi- poor, patient's positive rate 50%, Healthy Human Serum negative rate 98%.
1: 10,1: 100,1: 1000,1: 10000,1: 100000,5 are carried out to the positive sample of 1 part of 200ng/ml or more
Gradient is diluted, and is positively correlated through detection luminous intensity with concentration, Fitted logistic curves, R > 0.9.
Embodiment 4:
A kind of 1 chemoluminescence method kit of people's aquaporin, including sample treatment solution, enzyme standard liquid, to be coated with capture anti-
Solid phase carrier, luminous substrate, the standard items of body.
Sample treatment solution ingredient: the CB of pH9.4~10.0 contains 0.1% thymolphthalein.
Enzyme standard liquid main component: Avidin marks horseradish peroxidase mutase and the anti-recombined human aquaporin of biotin labeling
1 monoclonal antibody monoclonal antibody of albumen combines the conjugate formed.
Be coated with the solid phase carrier of capture antibody: the magnetism of coating anti-1 polyclonal antibody of recombined human aquaporin of mouse is micro-
Ball.
Luminous substrate: A liquid is that luminol content is 0.02M, and the TE solution of pH=8.0, B liquid is that every 100mL solution contains lemon
The TE solution of lemon acid 0.282g, 0.2g urea peroxide pH=8.0.
Standard items: three parts of 1 antibody concentrations of aquaporin containing recombined human are respectively 2ng/ml, 20ng/ml, 200ng/ml
CB solution.
By detecting to 10 parts of patients and 100 parts of Healthy People volunteer's urines, mean value is detected plus 5 marks with volunteer
Quasi- poor, patient's positive rate 80%, Healthy Human Serum negative rate 100%.
1: 10,1: 100,1: 1000,1: 10000,1: 100000,5 are carried out to the positive sample of 1 part of 200ng/ml or more
Gradient is diluted, and is positively correlated through detection luminous intensity with concentration, Fitted logistic curves, R > 0.9.
Term used in this specification " antibody " refers to include chimeric or recombinant antibodies and monoclonal antibody and Duo Ke
Grand antibody or its proteolytic fragments, such as segment Fab or F (ab ') 2 etc..The method for generating antibody is those skilled in the art
It is well known, and be included in the prior art.
Term used in this specification " solid phase " includes common macromolecular organic plastics, nanoparticle, various tunica fibrosas
Deng and well known to a person skilled in the art for physics, chemistry, biological adsorption albumen material.
Claims (10)
1. a kind of 1 chemoluminescence method kit of people's aquaporin, including sample treatment solution, enzyme standard liquid, it is coated with capture antibody
Solid phase carrier, luminous substrate it is characterized by:
The capture antibody is anti-human 1 polyclonal antibody of aquaporin；
The enzyme standard liquid, main ingredient are that enzyme marks anti-human 1 monoclonal antibody of aquaporin.
2. the 1 chemoluminescence method kit of people's aquaporin according to benefit is wanted to require 1, it is characterised in that: the anti-human water
The immunogene of 1 antibody of channel protein is Purification of Human renal proximal tubular cell source aquaporin 1.
3. the 1 chemoluminescence method kit of people's aquaporin according to benefit is wanted to require 2, it is characterised in that: the Purification of Human
Renal proximal tubular cell source aquaporin 1 is 1 tetramer of Purification of Human renal proximal tubular cell source aquaporin.
4. the 1 chemoluminescence method kit of people's aquaporin according to benefit is wanted to require 1, it is characterised in that: the anti-human water
The immunogene of 1 antibody of channel protein is that can cross people's aquaporin 1 that eukaryotic expression cloned gene obtains.
5. the 1 chemoluminescence method kit of people's aquaporin according to claim 2 to 4, it is characterised in that: described anti-human
1 polyclonal antibody of aquaporin is one of source of mouse, Ma Yuan, Yang Yuan, rabbit or cavy source；The anti-human aquaporin egg
White 1 monoclonal antibody is one of source of mouse or rabbit source.
6. according to claim 1 to 4 it is one of any described in 1 chemoluminescence method kit of people's aquaporin, it is characterised in that:
The solid phase carrier is ELISA Plate or magnetic microsphere.
7. according to claim 1 to 4 it is one of any described in 1 chemoluminescence method kit of people's aquaporin, it is characterised in that:
The luminous substrate is that main component is luminol or its compound after modification as reducing agent and hydrogen peroxide, peroxidating
The peroxide agent of the horseradish peroxidases such as urea mutase sensitivity, reducing agent and oxidant can divide in same component or independently
Dress.
8. according to claim 1 to 4 it is one of any described in 1 chemoluminescence method kit of people's aquaporin, it is characterised in that:
The enzyme standard liquid is that Avidin marks horseradish peroxidase mutase and anti-human 1 monoclonal of aquaporin of biotin labeling anti-
Body monoclonal antibody combines the conjugate formed.
9. the 1 chemoluminescence method kit of people's aquaporin according to benefit is wanted to require 1, it is characterised in that: the sample
Treatment fluid main component is the CB of pH9.4~10.0 containing thymolphthalein.
10. the 1 chemoluminescence method kit of people's aquaporin according to benefit is wanted to require 1, it is characterised in that: the reagent
Box also contains 1 standard items of someone's aquaporin, and standard items quantity is no less than 3 parts, be respectively 1 concentration of people's aquaporin 2~
The CB solution of 200ng/ml.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114836541A (en) * | 2022-05-23 | 2022-08-02 | 中国药科大学 | Application of urine microvesicle protein as kidney cancer diagnosis marker |
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|---|---|---|---|---|
| US5741671A (en) * | 1991-12-12 | 1998-04-21 | The Johns Hopkins University | Isolation cloning and expression of transmembrane water channel aquaporin 1(AQP1) |
| CN103917869A (en) * | 2011-09-22 | 2014-07-09 | 洛斯安第斯大学 | Methods for monitoring, diagnosing and/or prognosing early acute kidney injury |
| CN106520933A (en) * | 2016-10-19 | 2017-03-22 | 佳木斯大学 | An expression positioning method for aquaporins in a human larynx tissue |
-
2017
- 2017-08-23 CN CN201710732285.1A patent/CN109425742A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5741671A (en) * | 1991-12-12 | 1998-04-21 | The Johns Hopkins University | Isolation cloning and expression of transmembrane water channel aquaporin 1(AQP1) |
| CN103917869A (en) * | 2011-09-22 | 2014-07-09 | 洛斯安第斯大学 | Methods for monitoring, diagnosing and/or prognosing early acute kidney injury |
| CN106520933A (en) * | 2016-10-19 | 2017-03-22 | 佳木斯大学 | An expression positioning method for aquaporins in a human larynx tissue |
Non-Patent Citations (2)
| Title |
|---|
| 上海信裕生物科技有限公司: "人水通道蛋白1（AQP1）酶联免疫分析试剂盒使用说明书", 《HTTP://WWW.BIOON.COM.CN/DOC/SHOWARTICLE.ASP?NEWSID=60508》 * |
| 李明堂 等: "猪水通道蛋白AQP1 多克隆抗体制备及其在猪体内的免疫定位", 《吉林农业大学学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114836541A (en) * | 2022-05-23 | 2022-08-02 | 中国药科大学 | Application of urine microvesicle protein as kidney cancer diagnosis marker |
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