CN102495218A - Enzyme-linked immunosorbent assay kit for 6-ketone-prostaglandin F1 alpha - Google Patents
Enzyme-linked immunosorbent assay kit for 6-ketone-prostaglandin F1 alpha Download PDFInfo
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- CN102495218A CN102495218A CN2011104200605A CN201110420060A CN102495218A CN 102495218 A CN102495218 A CN 102495218A CN 2011104200605 A CN2011104200605 A CN 2011104200605A CN 201110420060 A CN201110420060 A CN 201110420060A CN 102495218 A CN102495218 A CN 102495218A
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- ketone
- prostaglandin
- alpha
- pgf
- reagent kit
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Abstract
The invention discloses an enzyme-linked immunosorbent assay kit for 6-ketone-prostaglandin F1 alpha, which comprises a bovine serum albumin connection microtiter plate coated with the 6-ketone-PG F1 alpha, gelatin, a 6-ketone-PG F1 alpha standard substance, a 6-ketone-PG F1 alpha-IgG (immunoglobulin G) antibody, goat anti-rabbit IgG-horseradish peroxidase conjugate, o-phenylenediamine, diluent and substrate liquor. The enzyme-linked immunosorbent assay kit for the 6-ketone-prostaglandin F1 alpha has high specificity and can be used for effectively measuring the concentration of the 6-ketone-PG F1 alpha in blood plasma, blood cells, body fluids or tissues of humans and animals.
Description
Technical field
The invention belongs to ELISA measuring reagent kit, concrete a kind of 6-ketone-prostaglandin
ELISA measuring reagent kit that relates to.
Background technology
Prostaglandin is the primary product of vascular endothelial cell arachidonic acid metabolic, also can result from respiratory system, urinary system and digestive system etc.Prostaglandin is the strongest RA233 of body and vasodilator substance.
6-ketone-PGF
1 α(hereinafter to be referred as 6-ketone-PGF
1 α) be the stable metabolic product of prostacyclin.Blood plasma 6-ketone-PGF when diseases such as atherosclerotic, acute myocardial infarction, angina pectoris, cerebrovascular disease, diabetes, metastases, peripheral vessels thrombosis and thrombotic thrombocytopenic purpura
1 αContent often reduces.
Summary of the invention
The object of the present invention is to provide a kind of 6-ketone-prostaglandin
ELISA measuring reagent kit.
For realizing above-mentioned technical purpose, reach above-mentioned technique effect, the present invention realizes through following technical scheme:
6-ketone-prostaglandin of the present invention
ELISA measuring reagent kit comprises:
1) encapsulated 6-ketone-PGF
1 α-bovine serum albumin(BSA) connector ELISA Plate ,-20 ℃ of preservations;
2) gelatin;
3) 6-ketone-PGF
1 αStandard items, 4 ℃ of preservations;
4) antibody 6-ketone-PGF
1 α-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 (6-ketone-PGF
1 α-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 6-ketone-PGF from typical curve according to colour developing degree (OD value)
1 αContent.
Compared with prior art, the present invention has following beneficial effect:
1,6-ketone-prostaglandin of the present invention
ELISA measuring reagent kit closes and has good specificity, can effectively measure the concentration of blood plasma, haemocyte, body fluid or the 6-ketone-PGF1 in the tissue of humans and animals;
2, the antidiastole that is combined in the diseases such as atherosclerotic, acute myocardial infarction, angina pectoris, cerebrovascular disease, diabetes, metastases, peripheral vessels thrombosis and thrombotic thrombocytopenic purpura of 6-ketone-prostaglandin of the present invention
ELISA measuring reagent kit is highly sensitive, can guarantee good linear relationship;
3,6-ketone-prostaglandin of the present invention
ELISA 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 6-ketone-prostaglandin
ELISA measuring reagent kit, it comprises:
1) encapsulated 6-ketone-PGF
1 α-bovine serum albumin(BSA) connector ELISA Plate ,-20 ℃ of preservations;
2) gelatin, the 1g/ bottle;
3) 6-ketone-PGF
1 αStandard items, 1600pg/ml, 4 ℃ of preservations;
4) antibody 6-ketone-PGF
1 α-IgG, 1ml, titre is 1:8000,4 ℃ 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 the detection step of 6-ketone-prostaglandin of the present invention
ELISA measuring reagent kit following:
1, primary first-order equation
With 6-ketone-PGF
1 αStandard 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 6-ketone-PGF with 0.1% gelatin (PBS preparation)
1 α-IgG does the 1:10 dilution.
Zero standard (B0) hole adds dilution 100ul, and standard (ST) hole adds 6-ketone-PGF
1 αStandard items 100ul, sample (SMP) hole adds testing sample 100ul.
Each Kong Jun adds antibody 6-ketone-PGF
1 α-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 6-ketone-PGF
1 αStandard 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 6-ketone-PGF from typical curve
1 αConcentration.
[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 6-ketone-PGF in the blood plasma of mensuration humans and animals, haemocyte, body fluid or the tissue
1 αConcentration, 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. 6-ketone-prostaglandin
ELISA measuring reagent kit; It is characterized in that, comprising:
1) encapsulated 6-ketone-PGF
1 α-bovine serum albumin(BSA) connector ELISA Plate;
2) gelatin;
3) 6-ketone-PGF
1 αStandard items;
4) antibody 6-ketone-PGF
1 α-IgG;
5) goat anti-rabbit igg-horseradish peroxidase bond;
6) o-phenylenediamine;
7) dilution;
8) substrate solution.
2. 6-ketone-prostaglandin according to claim 1
ELISA measuring reagent kit, it is characterized in that: said gelatin is the 1g/ bottle.
3. 6-ketone-prostaglandin according to claim 1
ELISA measuring reagent kit is characterized in that: said 6-ketone-PGF
1Standard items are 1600pg/ml.
4. 6-ketone-prostaglandin according to claim 1
ELISA measuring reagent kit is characterized in that: said antibody 6-ketone-PGF
1 α-IgG is 1ml, and titre is 1:8000.
5. 6-ketone-prostaglandin according to claim 1
ELISA measuring reagent kit, it is characterized in that: said goat anti-rabbit igg-horseradish peroxidase bond is 100ug/ml.
6. 6-ketone-prostaglandin according to claim 1
ELISA measuring reagent kit, it is characterized in that: said o-phenylenediamine is the 15mg/ bottle.
7. 6-ketone-prostaglandin according to claim 1
ELISA measuring reagent kit, it is characterized in that: dilution is 0.2M.
8. according to any described 6-ketone-prostaglandin
ELISA measuring reagent kit in the claim 1 to 7, it is characterized in that: substrate solution is 0.1M.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011104200605A CN102495218A (en) | 2011-12-15 | 2011-12-15 | Enzyme-linked immunosorbent assay kit for 6-ketone-prostaglandin F1 alpha |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011104200605A CN102495218A (en) | 2011-12-15 | 2011-12-15 | Enzyme-linked immunosorbent assay kit for 6-ketone-prostaglandin F1 alpha |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102495218A true CN102495218A (en) | 2012-06-13 |
Family
ID=46187059
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011104200605A Pending CN102495218A (en) | 2011-12-15 | 2011-12-15 | Enzyme-linked immunosorbent assay kit for 6-ketone-prostaglandin F1 alpha |
Country Status (1)
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1995009645A1 (en) * | 1993-10-07 | 1995-04-13 | The George Washington University Medical Center | Method of treating septic shock by preventing actin polymerization |
| CN102086228A (en) * | 2009-12-04 | 2011-06-08 | 北京维德维康生物技术有限公司 | ELISA (Enzyme Linked Immunosorbent Assay) Kit and application thereof |
| CN201993363U (en) * | 2010-10-27 | 2011-09-28 | 北京勤邦生物技术有限公司 | Lincomycin enzyme-linked immunosorbent assay (ELISA) detection kit |
| CN102234333A (en) * | 2010-05-08 | 2011-11-09 | 国家海洋环境监测中心 | Establishment of preparation and rapid detection method for polyclonal antiserum of DDT (dichlorodiphenyltrichloroethane) metabolite DDA (2,2-bis(4-chlorophenyl)acetic acid) |
-
2011
- 2011-12-15 CN CN2011104200605A patent/CN102495218A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1995009645A1 (en) * | 1993-10-07 | 1995-04-13 | The George Washington University Medical Center | Method of treating septic shock by preventing actin polymerization |
| CN102086228A (en) * | 2009-12-04 | 2011-06-08 | 北京维德维康生物技术有限公司 | ELISA (Enzyme Linked Immunosorbent Assay) Kit and application thereof |
| CN102234333A (en) * | 2010-05-08 | 2011-11-09 | 国家海洋环境监测中心 | Establishment of preparation and rapid detection method for polyclonal antiserum of DDT (dichlorodiphenyltrichloroethane) metabolite DDA (2,2-bis(4-chlorophenyl)acetic acid) |
| CN201993363U (en) * | 2010-10-27 | 2011-09-28 | 北京勤邦生物技术有限公司 | Lincomycin enzyme-linked immunosorbent assay (ELISA) detection kit |
Non-Patent Citations (4)
| Title |
|---|
| 《中国中西医结合急救杂志》 20101130 易小民等 全蝎纯化液对凝血酶诱导血管内皮细胞释放血管性血友病因子和6-酮-前列腺素F1alpha的影响 334-336 1-8 第17卷, 第6期 * |
| 卢国元等: "狼疮肾炎患者血浆花生四烯酸代谢新指标的变化", 《中华风湿病学杂志》 * |
| 卢国元等: "血浆花生四烯酸代谢新指标在糖尿病肾病患者中的变化及意义", 《江苏医药》 * |
| 易小民等: "全蝎纯化液对凝血酶诱导血管内皮细胞释放血管性血友病因子和6-酮-前列腺素F1α的影响", 《中国中西医结合急救杂志》 * |
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Application publication date: 20120613 |