CN102495219A - Prostaglandin E2 enzyme-linked immunoassay kit - Google Patents
Prostaglandin E2 enzyme-linked immunoassay kit Download PDFInfo
- Publication number
- CN102495219A CN102495219A CN2011104200732A CN201110420073A CN102495219A CN 102495219 A CN102495219 A CN 102495219A CN 2011104200732 A CN2011104200732 A CN 2011104200732A CN 201110420073 A CN201110420073 A CN 201110420073A CN 102495219 A CN102495219 A CN 102495219A
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- CN
- China
- Prior art keywords
- pge
- pge2
- reagent kit
- measuring reagent
- enzyme
- Prior art date
- 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.)
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Abstract
The invention discloses a prostaglandin E2 (PGE2) enzyme-linked immunoassay kit, which comprises PGE2-coated bovine serum albumin junctional complex enzyme label, gelatin, a PGE2 standard substance, an antibody PGE2-IgG, a goat anti rabbit IgG-horseradish peroxidase enzyme bonding substance, o-phenylenediamine, diluent and a substrate solution. The prostaglandin E2 (PGE2) enzyme-linked immunoassay kit has high specificity, and can effectively measure the PGE2 concentration of blood plasma, blood cells, body fluid or tissues of human and animals.
Description
Technical field
The invention belongs to ELISA measuring reagent kit, concrete relate to a kind of PGE
2ELISA measuring reagent kit.
Background technology
PGE
2(hereinafter to be referred as PGE
2) be a lot of tissues of body and the arachidonic primary product of organ (like nerve, intestines and stomach, respiratory tract and uropoiesis, reproductive system).Measure the PGE of blood, gastric juice, cerebrospinal fluid, urine, seminal fluid or various tissues
2Content back mirrors the function of relevant organ or tissue.
Summary of the invention
The object of the present invention is to provide a kind of PGE
2ELISA measuring reagent kit.
For realizing above-mentioned technical purpose, reach above-mentioned technique effect, the present invention realizes through following technical scheme:
PGE of the present invention
2ELISA measuring reagent kit comprises:
1) encapsulated PGE
2-bovine serum albumin(BSA) connector ELISA Plate ,-20 ℃ of preservations;
2) gelatin;
3) PGE
2Standard items, 4 ℃ of preservations;
4) antibody PGE
2-IgG, 4 ℃ of preservations;
5) goat anti-rabbit igg-horseradish peroxidase bond ,-20 ℃ of preservations;
6) o-phenylenediamine;
7) dilution is done the 1:20 dilution with preceding with distilled water;
8) substrate solution is done the 1:5 dilution with preceding with distilled water.
Detection principle of the present invention is following:
Antigen (PGE
2-BSA) encapsulate solid phase carrier, with competitive ground of free antigen (standard items or testing sample) and a certain amount of antibodies that gets, add excessive enzyme labelled antibody after the washing, add the substrate colour developing again.Can extrapolate testing sample PGE from typical curve according to colour developing degree (OD value)
2Content.
Compared with prior art, the present invention has following beneficial effect:
1, PGE of the present invention
2ELISA measuring reagent kit closes has good specificity, can effectively measure the PGE in blood, gastric juice, cerebrospinal fluid, urine, seminal fluid or the various tissue
2Concentration;
2, PGE of the present invention
2The antidiastole that ELISA measuring reagent kit is combined in the diseases such as the nervous system disease, asthma, peptic ulcer, disease in the urological system, sterility and tumour is highly sensitive, can guarantee good linear relationship;
3, PGE of the present invention
2ELISA measuring reagent kit is combined under-20 ℃ of conditions cocoa and preserved 6 months.
Above-mentioned explanation only is the general introduction of technical scheme of the present invention, understands technological technical scheme of the present invention in order can more to know, and can implement according to the content of instructions, below with preferred embodiment of the present invention.
Embodiment
A kind of PGE
2ELISA measuring reagent kit, it comprises:
1) encapsulated PGE
2-bovine serum albumin(BSA) connector ELISA Plate ,-20 ℃ of preservations;
2) gelatin, the 1g/ bottle;
3) PGE
2Standard items, 1600pg/ml, 4 ℃ of preservations;
4) antibody PGE
2-IgG, 1ml, titre is 1:2000,4 ℃ of preservations;
5) goat anti-rabbit igg-horseradish peroxidase bond (GARIgG-HRP), 100ug/ml ,-20 ℃ of preservations;
6) o-phenylenediamine (OPD), the 15mg/ bottle;
7) dilution (PBS), 0.2M does the 1:20 dilution with preceding with distilled water;
8) substrate solution (Buffer2), 0.1M does the 1:5 dilution with preceding with distilled water.
Utilize PGE of the present invention
2The detection step of ELISA measuring reagent kit is following:
1, primary first-order equation
With PGE
2Standard items become 8 concentration with 0.1% gelatin doubling dilution: 1.25,2.5,5,10,20,40,80 and 160 pg/0.1ml will resist PGE with 0.1% gelatin (PBS preparation)
2-IgG does the 1:10 dilution.
Zero standard (B0) hole adds dilution 100ul, and standard (ST) hole adds PGE
2Standard items 100ul, sample (SMP) hole adds testing sample 100ul.
Each Kong Jun adds antibody PGE
2-IgG100ul (except the TA hole).
4hr under 37 ℃ of 2hr or the room temperature.
The tween-PBS dilution is washed plate 5 times.
2, secondary reaction
12ml dilutes GARIgG-HRP with 0.1% gelatin (diluent preparing), and each Kong Jun adds 200ul (except the TA hole).4hr under 37 ℃ of 2hr or the room temperature.
Tween-PBS washes plate 5 times.
3, reaction assay
With substrate solution fresh OPD-H
2O
2(OPD concentration 1mg/ml adds 0.5 microlitre, 30% H to matrix in every milliliter of substrate solution
2O
2), 200ul is injected in every hole.
Place 20min or to developing the color when better, with 3M sulfuric acid 50ul cessation reaction for 37 ℃.
Put room temperature 10min, on enzyme mark detector, measure OD
490, return to zero with the TA hole.
4, calculate
With PGE
2Standard items content is horizontal ordinate, corresponding B/B
0The % value is an ordinate, on semilogarithmic paper, makes typical curve, with sample well B/B
0The % value is calculated its PGE from typical curve
2Concentration.
[collection of specimens and preservation]: use 2%EDTA-Na
2(containing 0.7%NaCl) 1:9 anti-freezing, blood collection procedure need no bubble smoothly, prevent platelet activation, and urine specimen is left and taken and need be put antiseptic, and sample all needs 3000rpm/min immediately, behind the centrifugal 15min, gets blood plasma and sample to be measured, freezes-20 ℃, can preserve 3 months.
[the reagent scope of application and the term of validity]: the PGE in the blood plasma of mensuration humans and animals, haemocyte, body fluid or the tissue
2Concentration, this medicine box term of validity can be preserved 6 months (20 ℃).
The above is merely preferred case study on implementation of the present invention, is not limited to the present invention, and for a person skilled in the art, the present invention can have various changes and variation.All within spirit of the present invention and principle, any modification of being done, be equal to replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (8)
1. PGE
2ELISA measuring reagent kit is characterized in that, comprising:
1) encapsulated PGE
2-bovine serum albumin(BSA) connector ELISA Plate;
2) gelatin;
3) PGE
2Standard items;
4) antibody PGE
2-IgG;
5) goat anti-rabbit igg-horseradish peroxidase bond;
6) o-phenylenediamine;
7) dilution;
8) substrate solution.
2. PGE according to claim 1
2ELISA measuring reagent kit is characterized in that: said gelatin is the 1g/ bottle.
3. PGE according to claim 1
2ELISA measuring reagent kit is characterized in that: said PGE
2Standard items are 1600pg/ml.
4. PGE according to claim 1
2ELISA measuring reagent kit is characterized in that: said antibody PGE
2-IgG is 1ml, and titre is 1:2000.
5. PGE according to claim 1
2ELISA measuring reagent kit is characterized in that: goat anti-rabbit igg-horseradish peroxidase bond is 100ug/ml.
6. PGE according to claim 1
2ELISA measuring reagent kit is characterized in that: said o-phenylenediamine is the 15mg/ bottle.
7. PGE according to claim 1
2ELISA measuring reagent kit is characterized in that: dilution is 0.2M.
8. according to any PGE in the claim 1 to 7
2ELISA measuring reagent kit is characterized in that: substrate solution is 0.1M.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011104200732A CN102495219A (en) | 2011-12-15 | 2011-12-15 | Prostaglandin E2 enzyme-linked immunoassay kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011104200732A CN102495219A (en) | 2011-12-15 | 2011-12-15 | Prostaglandin E2 enzyme-linked immunoassay kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102495219A true CN102495219A (en) | 2012-06-13 |
Family
ID=46187060
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011104200732A Pending CN102495219A (en) | 2011-12-15 | 2011-12-15 | Prostaglandin E2 enzyme-linked immunoassay kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102495219A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105492906A (en) * | 2013-06-14 | 2016-04-13 | 首尔大学校产学协力团 | Method for detecting hypoxia or diagnosing hypoxia-related diseases |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006012259A1 (en) * | 2004-06-30 | 2006-02-02 | Aventis Pharmaceuticals Inc. | Methods involving micromosal prostaglandin e2 synthase |
| CN101949924A (en) * | 2010-09-29 | 2011-01-19 | 江南大学 | Enzyme-linked immunoassay method for benzophenone |
| CN102143760A (en) * | 2008-07-08 | 2011-08-03 | 雅培制药有限公司 | Prostaglandin E2 binding proteins and uses thereof |
-
2011
- 2011-12-15 CN CN2011104200732A patent/CN102495219A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006012259A1 (en) * | 2004-06-30 | 2006-02-02 | Aventis Pharmaceuticals Inc. | Methods involving micromosal prostaglandin e2 synthase |
| CN102143760A (en) * | 2008-07-08 | 2011-08-03 | 雅培制药有限公司 | Prostaglandin E2 binding proteins and uses thereof |
| CN101949924A (en) * | 2010-09-29 | 2011-01-19 | 江南大学 | Enzyme-linked immunoassay method for benzophenone |
Non-Patent Citations (1)
| Title |
|---|
| 何杨等: "酶联前列腺素E2方法的建立及临床应用", 《实验血液学杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105492906A (en) * | 2013-06-14 | 2016-04-13 | 首尔大学校产学协力团 | Method for detecting hypoxia or diagnosing hypoxia-related diseases |
| CN105492906B (en) * | 2013-06-14 | 2017-08-25 | 首尔大学校产学协力团 | Purposes of the arachidonic acid or derivatives thereof in detection obstructive sleep apnea |
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| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
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Application publication date: 20120613 |