CN205473834U - Simple and easy high flux thioredoxin activity detection kit - Google Patents
Simple and easy high flux thioredoxin activity detection kit Download PDFInfo
- Publication number
- CN205473834U CN205473834U CN201620150746.5U CN201620150746U CN205473834U CN 205473834 U CN205473834 U CN 205473834U CN 201620150746 U CN201620150746 U CN 201620150746U CN 205473834 U CN205473834 U CN 205473834U
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- China
- Prior art keywords
- kit
- buffer
- thioredoxin
- high flux
- detection kit
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
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- 108060008226 thioredoxin Proteins 0.000 title claims abstract description 35
- 102000002933 Thioredoxin Human genes 0.000 title claims abstract description 29
- 238000001514 detection method Methods 0.000 title claims abstract description 29
- 229940094937 thioredoxin Drugs 0.000 title claims abstract description 29
- 230000004907 flux Effects 0.000 title claims abstract description 16
- 230000000694 effects Effects 0.000 title claims abstract description 7
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 22
- 108010071390 Serum Albumin Proteins 0.000 claims abstract description 9
- 102000007562 Serum Albumin Human genes 0.000 claims abstract description 9
- 239000000758 substrate Substances 0.000 claims abstract description 9
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical group C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 claims abstract description 8
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000007788 liquid Substances 0.000 claims abstract description 8
- 239000000203 mixture Substances 0.000 claims abstract description 4
- 239000000872 buffer Substances 0.000 claims description 26
- 238000012360 testing method Methods 0.000 claims description 16
- 102100036407 Thioredoxin Human genes 0.000 claims description 6
- 230000002255 enzymatic effect Effects 0.000 claims description 6
- 108091006905 Human Serum Albumin Proteins 0.000 claims description 3
- 102000008100 Human Serum Albumin Human genes 0.000 claims description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 2
- 229940098773 bovine serum albumin Drugs 0.000 claims description 2
- 239000000243 solution Substances 0.000 abstract description 9
- 239000007853 buffer solution Substances 0.000 abstract description 8
- 108090000790 Enzymes Proteins 0.000 abstract description 3
- 102000004190 Enzymes Human genes 0.000 abstract description 3
- ACFIXJIJDZMPPO-NNYOXOHSSA-N NADPH Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](OP(O)(O)=O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 ACFIXJIJDZMPPO-NNYOXOHSSA-N 0.000 abstract description 3
- 229960000789 guanidine hydrochloride Drugs 0.000 abstract description 3
- 238000007689 inspection Methods 0.000 abstract description 3
- 239000006166 lysate Substances 0.000 abstract description 3
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 abstract description 3
- 238000000746 purification Methods 0.000 abstract description 3
- 238000004458 analytical method Methods 0.000 abstract description 2
- 239000008057 potassium phosphate buffer Substances 0.000 abstract 1
- 238000000034 method Methods 0.000 description 8
- 210000001519 tissue Anatomy 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 4
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 239000003085 diluting agent Substances 0.000 description 3
- 238000008157 ELISA kit Methods 0.000 description 2
- 102000004877 Insulin Human genes 0.000 description 2
- 108090001061 Insulin Proteins 0.000 description 2
- 238000007630 basic procedure Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 229940125396 insulin Drugs 0.000 description 2
- 210000002381 plasma Anatomy 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 101100128227 Caenorhabditis elegans lid-1 gene Proteins 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 101000852559 Homo sapiens Thioredoxin Proteins 0.000 description 1
- 102000013090 Thioredoxin-Disulfide Reductase Human genes 0.000 description 1
- 108010079911 Thioredoxin-disulfide reductase Proteins 0.000 description 1
- XOCUXOWLYLLJLV-UHFFFAOYSA-N [O].[S] Chemical compound [O].[S] XOCUXOWLYLLJLV-UHFFFAOYSA-N 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000003722 extracellular fluid Anatomy 0.000 description 1
- 108700022423 fluorescein-isothiocyanated- insulin Proteins 0.000 description 1
- 102000056461 human TXN Human genes 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The utility model belongs to the technical field of analysis and detection, involve a simple and easy high flux thioredoxin activity detection kit. Four reagent bottles in it contains reagent lid, kit and places the kit in, four reagent bottles respectively splendid attires detect buffer solutions, terminate buffer solution, enzyme solution and standard liquid, the detection buffer solution is NADPH, substrate and potassium phosphate buffer solution, it is DTNB and guanidine hydrochloride solution to terminate the buffer solution, the standard liquid is 5 mu gmL's people thioredoxin and the mixed liquid of 0.1%wt serum albumin. Compared with the prior art, the utility model discloses detectivity is high, meanwhile, the combination configuration of its composition can satisfy the needs of the basic flow configuration of existing commonly used automated inspection equipment basically. In addition, this kit can also detect the cell, organize lysate and purification thioredoxin active, and does not have unique tissue specificity.
Description
Technical field
This utility model belongs to technical field of analysis and detection, it particularly relates to a kind of easy high flux sulfur oxygen also
Protein active detection kit.
Background technology
Thioredoxin is present in all living cells, closely related with survival with cell growth.Thioredoxin also can
It is secreted into extracellular, is present in blood and extracellular fluid.The change of this protein active is relevant with the generation development of numerous disease.Mesh
Before, thioredoxin ELISA kit, is based on immunological response, relies on antigen, the specific reaction of antibody, detection
The protein level of thioredoxin.But, thioredoxin is easily modified change activity by internal metabolite, intermediate product,
The protein level of thioredoxin in tissue and cell, it is impossible to reflect the functional status of thioredoxin definitely.Additionally,
ELISA kit has the tissue specificity of uniqueness, may use the sample of separate sources, need to distinguish which kind of ELISA in experiment
Test kit can survey the sample in hands.So far, in laboratory research, the measuring method for activity of thioredoxin mainly has two kinds: insulin
Survey method (Luthman, the Biochemistry 21:6628-6633 that lives；1982.), end-point method (Arner, Methods Enzymol
300:226-239.；1999.) end-point method (Wu, the Y.Atherosclerosis 212:351-355 and after improvement；2010.).
Insulin surveys the method for living, and is suitable for enzyme dynamics, is not suitable for the mensuration of high-volume sample.End-point method after end-point method and improvement,
Condition and standard used by each laboratory are not quite similar, and step is more complicated, and measured result is difficult to compare.
Summary of the invention
For solving above-mentioned technical problem, this utility model provides a kind of high sensitivity, efficient and easy to use simple
High flux thioredoxin active detection kit.
One described in the utility model easy high flux thioredoxin active detection kit, described Activity determination
Test kit comprises reagent lid 1, test kit 3 and four reagent bottle 2 being placed in test kit；Described four reagent bottle 2 points
Sheng Zhuan not detect buffer, stop buffer, enzymatic solution and titer；Described detection buffer is NADPH-substrate buffer solution；
Described stop buffer is the guanidinium-hydrochloride buffer of DTNB；Described titer is the serum albumin dilution of the Trx
Liquid.
Described NADPH-substrate buffer solution is NADPH, substrate and kaliumphosphate buffer；The guanidinium-hydrochloride buffer of described DTNB
For DTNB and guanidine hydrochloride solution；The serum albumin diluent of the described Trx is the Trx of 5 μ g/mL
With 0.1%wt serum albumin mixed liquor.
Described reagent bottle 2 is plastic bottle or EP pipe.
Described serum albumin is human serum albumin or bovine serum albumin.
Described reagent lid 1 buckle closure constitutes the cube that outside size is 10cm × 10cm × 3cm on test kit 3.
The capacity of the reagent bottle 2 of above-mentioned splendid attire detection buffer, stop buffer, enzymatic solution and titer is respectively
3.7mL, 15mL, 100 μ L and 200 μ L.
Compared with prior art, easy high flux thioredoxin active detection kit described in the utility model inspection
Survey highly sensitive, it is sufficient to tell the signal that in 20 microlitre human serum/blood plasma, thioredoxin provides；Meanwhile, its composition
Combination configuration can substantially meet existing conventional automatic checkout equipment basic procedure configuration needs.Additionally, this test kit is also
Cell, Tissue lysates and purification thioredoxin active can be detected, and without unique tissue specificity.
Accompanying drawing explanation
Fig. 1: easy high flux thioredoxin active detection kit component diagram；Reagent lid-1, reagent bottle-
2, test kit-3.
Detailed description of the invention
Below in conjunction with specific embodiment, easy high flux thioredoxin active described in the utility model is detected
Test kit is described further, but protection domain of the present utility model is not limited to this.
Embodiment 1
A kind of easy high flux thioredoxin active detection kit, described activity detection kit comprises test kit
Lid 1, test kit 3 and four reagent bottle 2 being placed in test kit；Four described reagent bottle 2 respectively contain detection buffer,
Stop buffer, enzymatic solution and titer；Described detection buffer is NADPH-substrate buffer solution；Described stop buffer is
The guanidinium-hydrochloride buffer of DTNB；Described titer is the serum albumin diluent of the Trx.
Described NADPH-substrate buffer solution is 2mM NADPH, 10 μMs of substrates and 250mM kaliumphosphate buffer (pH=7.5),
Substrate is FITC-insulin；The guanidinium-hydrochloride buffer of described DTNB is 1mM DTNB and 8M guanidine hydrochloride solution；Described human thioredoxin is also
The serum albumin diluent of albumen is the Trx (hTrx1) and the 0.1%wt human serum albumin mixing of 5 μ g/mL
Liquid；Described enzymatic solution is 5 μMs of cattle thioredoxin reductases.Described reagent bottle 2 is managed for EP.Described test kit lid 1 and reagent
Box frame 3 can constitute the cube that outside size is 10cm × 10cm × 3cm.Described detection buffer, stop buffer, enzyme are molten
Capacity and the splendid attire capacity of the reagent bottle 2 corresponding to liquid and titer are followed successively by 3.7mL, 15mL, 100 μ L and 200 μ L.
Compared with prior art, easy high flux thioredoxin active detection kit described in the utility model inspection
Survey highly sensitive, it is sufficient to tell the signal that in 20 microlitre human serum/blood plasma, thioredoxin provides；Meanwhile, its composition
Combination configuration can substantially meet existing conventional automatic checkout equipment basic procedure configuration needs.Additionally, this test kit is also
Cell, Tissue lysates and purification thioredoxin active can be detected, and without unique tissue specificity.
Claims (5)
1. an easy high flux thioredoxin active detection kit, it is characterised in that described activity detection kit
Comprise reagent lid (1), test kit (3) and four reagent bottle (2) being placed in test kit；Four described reagent bottle (2)
Contain detection buffer, stop buffer, enzymatic solution and titer respectively；Described detection buffer is NADPH-substrate buffer
Liquid；Described stop buffer is the guanidinium-hydrochloride buffer of DTNB；Described titer is that the serum albumin of the Trx is dilute
Release liquid.
One the most according to claim 1 easy high flux thioredoxin active detection kit, it is characterised in that
Described reagent bottle (2) is plastic bottle or EP pipe.
One the most according to claim 1 easy high flux thioredoxin active detection kit, it is characterised in that
Described serum albumin is human serum albumin or bovine serum albumin.
One the most according to claim 1 easy high flux thioredoxin active detection kit, it is characterised in that
Described reagent lid (1) buckle closure is at the cube that test kit (3) the outside size of upper composition is 10cm × 10cm × 3cm.
One the most according to claim 1 easy high flux thioredoxin active detection kit, it is characterised in that
The capacity of the reagent bottle (2) containing detection buffer, stop buffer, enzymatic solution and titer be respectively 3.7mL, 15mL, 100
μ L and 200 μ L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201620150746.5U CN205473834U (en) | 2016-02-29 | 2016-02-29 | Simple and easy high flux thioredoxin activity detection kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201620150746.5U CN205473834U (en) | 2016-02-29 | 2016-02-29 | Simple and easy high flux thioredoxin activity detection kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN205473834U true CN205473834U (en) | 2016-08-17 |
Family
ID=56679386
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201620150746.5U Expired - Fee Related CN205473834U (en) | 2016-02-29 | 2016-02-29 | Simple and easy high flux thioredoxin activity detection kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN205473834U (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108627469A (en) * | 2017-03-21 | 2018-10-09 | 凯熙医药（武汉）股份有限公司 | A kind of thioredoxin reductase activity test method for cooperation detection equipment |
-
2016
- 2016-02-29 CN CN201620150746.5U patent/CN205473834U/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108627469A (en) * | 2017-03-21 | 2018-10-09 | 凯熙医药（武汉）股份有限公司 | A kind of thioredoxin reductase activity test method for cooperation detection equipment |
| CN108627469B (en) * | 2017-03-21 | 2021-04-02 | 凯熙医药（武汉）股份有限公司 | Thioredoxin reductase activity detection method for cooperative detection equipment |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20220818 Address after: 19 Yuquan Road (a), Shijingshan District, Beijing 100049 Patentee after: University OF CHINESE ACADEMY OF SCIENCES Address before: 100190 In the Youth Apartment Park, University of Chinese Academy of Sciences, No. 80, Zhongguancun East Road, Haidian District, Beijing 100190 Patentee before: CHINA SCIENCES RESEARCH SOURCE SCIENCE & TECHNOLOGY DEVELOPMENT Co.,Ltd. |
|
| TR01 | Transfer of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160817 |
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| CF01 | Termination of patent right due to non-payment of annual fee |