CN1381725A - Process for preparing diagnosing reagent kit of alkaline phosphatase and isozyme - Google Patents
Process for preparing diagnosing reagent kit of alkaline phosphatase and isozyme Download PDFInfo
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- CN1381725A CN1381725A CN 01110756 CN01110756A CN1381725A CN 1381725 A CN1381725 A CN 1381725A CN 01110756 CN01110756 CN 01110756 CN 01110756 A CN01110756 A CN 01110756A CN 1381725 A CN1381725 A CN 1381725A
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Abstract
A diagnosing reagent kit of alkaline phosphotase isozyme (a hydrolase) and its preparing process are disclosed. It features that its reagents are respectively loaded in different ampules. Its advantages are high stability and repeatability, high speed and simple operation.
Description
The present invention relates to a kind of hydrolytic enzyme---the preparation of alkaline phosphatase isoenzyme diagnostic kit belongs to the zymetology field.
In recent years, zymetology is very fast with the application development in the treatment in clinical diagnosis, now develops into two frontiers in the medical science respectively, promptly diagnoses zymetology and treatment zymetology.At present existing hundreds of enzyme test can be used for clinical diagnosis.This development of clinical enzymology, part are because the development of determination techniques makes the mensuration of many enzymes accurate and easy day by day, can be used for conventional determining.Part is owing to recognize in some disease progression process enzyme activity this objective fact that must change, early stage in disease particularly, and when also failing to find focus with X-ray sheet, CT, and the interior enzymatic activity of body has begun to change.If can regularly have a medical check-up, can accomplish early detection to disease.What deserves to be mentioned is that the application majority of diagnosing zymetology at present clinically is the mensuration to the gross activity of enzyme, and existing corresponding reagent box is sold on the market, its shortcoming is to lack organ specificity.The kit of at present relevant enzyme assay has 20～30 kinds approximately, and the isodynamic enzyme detection kit has 3～5 kinds approximately, but most usefulness is Chemical Inhibition Method or immunodepression.And isodynamic enzyme is meant that the zymoprotein structure is different and have the class of enzymes of identical catalysis characteristics, and different isodynamic enzymes is from different tissues and cell.When a certain tissue or organ generation pathology, because the variation of tissue damaged or permeability of cell membrane, cause that desmoenzyme is released into blood in a large number and causes the change of sero-enzyme activity.Detection by isodynamic enzyme can be determined disease sites, thereby improves the specificity and the susceptibility of zymetology diagnosis, but judging prognosis also.At present show liquid owing to prepare enzyme voluntarily, because of agents useful for same various in style, diversified in specifications, cause enzyme display result instability, the result judges disunity, and the isodynamic enzyme detecting operation is more loaded down with trivial details, time-consuming or the like, the particularly application of large-scale Biochemical Analyzer, the operation that clinical biochemical is detected is easier, saves time, laborsaving, but it is the mensuration to total enzyme activity, inorganizable specificity is compared, and just accurately how the detection of isodynamic enzyme make the early stage qualitative and location of pathology.
The object of the present invention is to provide a kind of superior in quality, stable performance, repetition performance good, simplified simultaneously the operating process that detects greatly and used this kit can make the detecting operation standardization of isodynamic enzyme, thereby made the detection of enzymatic activity and quality control reach the preparation of the hydrolytic enzyme diagnostic kit of unified standard.
Purpose of the present invention is implemented by following technical scheme:
Do cellulose acetate electrophoresis, it includes: (1), compound concentration are 0.02～0.1mol/L electrode solution, and pH is 7.5-9.0; (2), use glacial acetic acid as fixing agent; (3), compound concentration is the fine film damping fluid of 0.03～0.2mol/L bubble vinegar; In the fine film damping fluid of (3) described bubble vinegar packing ampoule A; With (2) described fixing agent packing ampoule E; (4) preparation colour developing liquid
(a), compound concentration is 0.05～0.3mol/L damping fluid, pH is 9.0～11.0, among the packing ampoule B,
(b) with substrate and diazol colours packing ampoule C;
(c), with agarose or agar packing ampoule D; Content agarose or the agar among content substrate among the ampoule C and diazol colours, the ampoule D wherein, by weight percentage, substrate, diazol colours, agarose or agar are respectively 20.0%～86.7%, 10.5%～75.2%, 1.2%～28.6%, with concentration is that the damping fluid of 0.05～0.3mol/L is 100ml～250ml packing ampoule B, time spent adds the water heating for dissolving with ampoule D, and the contents mixed with ampoule B, ampoule C, ampoule D is colour developing liquid then.
Do agarose gel electrophoresis, it includes:
(1), compound concentration is 0.02～0.1mol/L electrode solution, pH is 7.5-9.0
(2), use glacial acetic acid as fixing agent packing ampoule E;
(3), the preparation of Ago-Gel: it is the fine film damping fluid of 0.05～0.2mol/L bubble vinegar 80ml～120ml that agarose (or agar) 0.5g-1g is added concentration, heating for dissolving, packing while hot is prepared as ampoule A; Or the preparation of Ago-Gel also can be with agarose (or agar), sucrose and polyvinylpyrrolidone, be respectively 7.0%～32.3%, 55.6%～91.0%, 1.4%～19.4% by weight percentage, adding 100ml～250ml concentration is the fine film damping fluid of bubble vinegar of 0.05mol/L～0.2mol/L, heating for dissolving, packing while hot is prepared as ampoule A.
(4) preparation colour developing liquid
(a), compound concentration is 0.05～0.3mol/L damping fluid, pH is 9.0～11.0, among the packing ampoule B,
(b) with substrate and diazol colours packing ampoule C;
(c), with agarose or agar packing ampoule D; Content agarose or the agar among content substrate among the ampoule C and diazol colours, the ampoule D wherein, by weight percentage, substrate, diazol colours, agarose or agar are respectively 20.0%～86.7%, 10.5%～75.2%, 1.2%～28.6%, with concentration is damping fluid 100ml～250ml packing ampoule B of 0.05～0.3mol/L, and the contents mixed of time spent with ampoule B, ampoule C, ampoule D is colour developing liquid.
Colour developing liquid can be prepared as freeze-dried powder reagent, and it is with dress damping fluid and substrate in the described ampoule B, the dissolving postlyophilization; It is that the 0.3g-1.5g substrate is added 100ml-250ml damping fluid packing ampoule B.Dress damping fluid and diazol colours in the described ampoule C, the dissolving postlyophilization, it is the damping fluid packing ampoule C that the diazol colours of 0.2g-1g is added 100ml-250ml; Time spent will be dissolved in water in the ampoule B, is dissolved in water in the ampoule C, with ampoule B, ampoule C contents mixed, is colour developing liquid.
The damping fluid of preparing among the described ampoule B, the damping fluid of the damping fluid of available Tris series, phosphoric acid series or borate buffer, diamido dimethyl 1.3 propylene glycol-HCL damping fluid;
Substrate can be with the phosphate of naphthols or its derivant among the described ampoule C, as alpha-Naphthol phosphoric acid or betanaphthol phosphoric acid or alpha-Naphthol phosphoric acid salt compound, naphthols AS-MX phosphoric acid, naphthols AS-BI phosphoric acid, or its naphthols AS-MX phosphoric acid salt and naphthols AS-BI phosphoric acid salt, toluidine-5-bromo-3-indoles phosphoric acid, nitrobenzene-5-bromo-3-indoles phosphoric acid, tricresyl phosphate amine salt; Diazol colours can with solid blue B or solid blue BB, solid blue R or blue RR, solid purple B, three-bristle cudrania garnet GBC, fast red RC, fast red TR, six azos be paid magenta etc. admittedly.
In addition, for improving specificity and the susceptibility of ALP, can be in conjunction with using inhibitor and heat inactivation method.In colour developing liquid, add following any reagent during colour developing, but specificity suppresses certain band and shows other composition.This all has important value for internal organs source of illustrating ALP and pathology location.
Specific inhibitor adds in colour developing liquid:
Concentration is 5～10mM L-phenylalanine, suppresses ALP4, ALP5;
Concentration is 5～10mM L-homoarginine, suppresses ALP2;
Concentration is 5～10mM L-tryptophane or leucine, suppresses ALP4;
Concentration is 3～5M urea, suppresses ALP3;
Concentration is the 1M ethylenediamine tetraacetic acid, and except that ALP4, all the other are all suppressed.
The heat inactivation method:
65 ℃ of water-baths 30 minutes, except that ALP4, all the other were all suppressed with sample.
Alkaline phosphatase isoenzyme (ALP EC 3.1.3.1) is the enzyme of the multiple phosphate of hydrolysis under alkali condition, is distributed widely in each tissue of human body.The detection of isodynamic enzyme helps the diagnosis and differential diagnosis of liver disease and bone illness.
Advantage of the present invention is: reagent is sub-packed in the ampoule, is convenient to preserve, and stable performance, good reproducibility has alleviated labour intensity, has changed the loaded down with trivial details of traditional artificial reagent preparation.And the difference that exists between individuality in the operating process that causes therefrom, thereby make regularized operation, make the detection of enzyme and quality control reach unified standard, use kit to simplify operating process simultaneously, save time, fast, be convenient to promote.Kit is more convenient during especially as a large amount of sample detection is suitable for, for the diagnosis and the antidiastole of clinical disease provides foundation.
Embodiment:
Embodiment 1, cellulose acetate electrophoresis, it includes: (1), compound concentration are electrode solution for the 0.02mol/L barbitol buffer solution, and pH is 8.0; (2), with glacial acetic acid as fixing agent, get 2ml packing ampoule E; (3), compound concentration is the fine film damping fluid of 0.2mol/L bubble vinegar, in the packing ampoule A; (4) preparation colour developing liquid: (a), compound concentration is 0.05mol/LTris damping fluid 100ml, pH is 9.0, among the packing ampoule B, (b) with substrate 3mg and diazol colours 10mg packing ampoule C; (c), with agarose 0.3mg packing ampoule D; After time spent adds the water heating for dissolving with content, the contents mixed of ampoule C, the ampoule D content of ampoule B, add the mixed liquor of content of content, the ampoule C of ampoule B, be colour developing liquid after the mixing.
Use: vacuolar membrane at first: open ampoule A and add water to 25ml, the fine film bubble of vinegar in this liquid 20 minutes.Then the fine film of vinegar is taken out, be clipped in middle suction of filter paper and remove excessive moisture, film bar frosted face up.With a print libation at an ancient wedding ceremony serum point at distance film one end 2cm place.The electrode buffer for preparing is poured in the electrophoresis tank, made the interior electrode buffer of groove be in same level.In the cathode and anode groove of electrophoresis tank respectively with 4 layers of both sides that rides over groove of folded filter paper of suitable dimension, as salt bridge.The film bar point sample of point sample is faced down, and the point sample place rides on the salt bridge towards cathode terminal, and the film bar needs to be adjacent to the exhibition of stretching tight with salt bridge.Energized, regulation voltage 100～260v, after 10～30 minutes, powered-down.Carry out the isodynamic enzyme colour developing then: liquid in the ampoule B is added ampoule C, make dissolving, pour dry powder in the ampoule D into a small test tube then, add 1ml water, heating for dissolving is then with ampoule
B, ampoule C mixed liquor add in the good ampoule D liquid of dissolving, and mixing is layered on the glass plate while hot, after the cooling, electrophoresis is crossed the film bar take out, and the point sample face is attached on the offset plate, 37 ℃ of incubations 20～30 minutes, and promptly visible isozymogram.
The result: serum alkaline phosphatase (ALP) isodynamic enzyme can occur ALP1-6 bar district band from positive pole to negative pole, and ALP2 (liver type) and two kinds of isodynamic enzymes of ALP3 (bone type) appear in normal human serum.When primary carcinoma of liver, cirrhosis, liver type activity of isoenzyme strengthens.Bone type activity of isoenzyme strengthens when osteogenic sarcoma, hyperthyroidism.In the gestation, ALP4 can appear in serum, sharply descends if ALP4 is active, usually is the dead diagnostic markers of placenta insufficiency or fetus, and other people ALP4 occurs, are shown with the tumour illness in advance, and ALP1 and ALP6 occur when pathologic condition.
Embodiment 2, do cellulose acetate electrophoresis, it includes: (1), compound concentration are electrode solution for the 0.1mol/Ltris-borate buffer, and pH is 9.0; (2), with glacial acetic acid as fixing agent, get 2.5ml packing ampoule E; (3), compound concentration is the fine film damping fluid of 0.1mol/Ltris-HCL bubble vinegar, in the packing ampoule A; (4) preparation colour developing liquid: (a), compound concentration is 0.2mol/L boric acid-NaOH damping fluid 100ml, pH is 10.0, among the packing ampoule B, (b) with substrate 10mg and diazol colours 6mg packing ampoule C; (c), with agarose 2mg packing ampoule D; After time spent adds the water heating for dissolving with content, the contents mixed of ampoule C, the ampoule D content of ampoule B, add the mixed liquor of content of content, the ampoule C of ampoule B, be colour developing liquid after the mixing.
Application is as described in the embodiment 1, and the result is also identical.
Embodiment 3, do agarose gel electrophoresis, it includes: (1), compound concentration are 0.08mol/Ltris-borate buffer electrode solution, and pH is 8.6; (2), with glacial acetic acid as fixing agent, get 2.5m1 packing ampoule E; (3), the preparation of Ago-Gel: it is the fine film damping fluid of 0.1mol/L bubble vinegar 80ml that agarose 0.5g is added concentration, heating for dissolving, packing while hot is prepared as ampoule A; (4) preparation colour developing liquid: (a), compound concentration is 0.2mol/L diamido dimethyl 1.3 propylene glycol damping fluid 100ml, pH is 11.0, among the packing ampoule B, (b) with substrate 15mg and diazol colours 6mg packing ampoule C; Time spent is colour developing liquid with the content of ampoule B, the contents mixed of ampoule C.
Use: with ampoule A water proof heating for dissolving, water while hot on a glass plate, the filter paper of 0.3 * 1cm is put in the cooling back at the about 2cm of distance film one end place.Take off filter paper after about 10 minutes,, placed 5 minutes at these place's application of sample 8 microlitres.Then gel slab being placed on the electrophoresis truss that has added electrode solution, puts up a bridge with 4 layers of gauze in two ends, voltage 100v, electrophoresis 30 minutes, powered-down.Offset plate take out is added the above-mentioned colour developing liquid that has mixed, be placed in 37 ℃ of incubators incubation 20-30 minute, promptly visible isodynamic enzyme colour developing band.
The result: serum alkaline phosphatase (ALP) isodynamic enzyme can occur ALP1-6 bar district band from positive pole to negative pole, and ALP2 (liver type) and two kinds of isodynamic enzymes of ALP3 (bone type) appear in normal human serum.When primary carcinoma of liver, cirrhosis, liver type activity of isoenzyme strengthens.Bone type activity of isoenzyme strengthens when osteogenic sarcoma, hyperthyroidism.In the gestation, ALP4 can appear in serum, sharply descends if ALP4 is active, usually is the dead diagnostic markers of placenta insufficiency or fetus, and other people ALP4 occurs, are shown with the tumour illness in advance, and ALP1 and ALP6 occur when pathologic condition.
Embodiment 4, do agarose gel electrophoresis, it includes: (1), compound concentration be the 0.03mol/L barbitol buffer solution as electrode solution, pH is 8.6; (2), with glacial acetic acid as fixing agent, get 2.5ml packing ampoule E; (3), the preparation of Ago-Gel is with agar 1g, sucrose 2g and polyvinylpyrrolidone 0.5g, adding 100ml concentration is the fine film damping fluid of bubble vinegar of 0.06mol/L, heating for dissolving, packing while hot is prepared as ampoule A.(4) preparation colour developing liquid: (a), compound concentration is 0.2mol/L boric acid-NaOH damping fluid 100ml, pH is 9.8, among the packing ampoule B, (b) with substrate 8mg and diazol colours 4mg packing ampoule C; The contents mixed of time spent with ampoule B, ampoule C is colour developing liquid.
Application is as described in the embodiment 3, and the result is also identical.
Embodiment 5, do cellulose acetate electrophoresis, it includes: (1), compound concentration are electrode solution for the 0.02mol/L barbitol buffer solution, and pH is 8.0; (2), with glacial acetic acid as fixing agent, get 2ml packing ampoule E; (3), compound concentration is the fine film damping fluid of 0.1mol/L bubble vinegar, in the packing ampoule A; (4) preparation colour developing liquid: (a), compound concentration is 0.05mol/LTris damping fluid 100ml, pH is 10.2, among the packing ampoule B, (b) with substrate 3mg and diazol colours 10mg packing ampoule F; (c), with agarose 0.3mg packing ampoule D; (d), with 6mmol/L L-phenylalanine packing ampoule E, after time spent adds the water heating for dissolving with contents mixed, the ampoule D content of the content of the content of ampoule B, ampoule C and ampoule E, the mixed liquor of the content of the content of adding ampoule B, the content of ampoule C, ampoule E, bed board while hot after the mixing is the colour developing offset plate.
Use: vacuolar membrane at first: open ampoule A and add water to 25ml, the fine film bubble of vinegar in this liquid 20 minutes.Then the fine film of vinegar is taken out, be clipped in middle suction of filter paper and remove excessive moisture, film bar frosted face up.With a print libation at an ancient wedding ceremony serum point at distance film one end 2cm place.The electrode buffer for preparing is poured in the electrophoresis tank, made the interior electrode buffer of groove be in same level.In the cathode and anode groove of electrophoresis tank respectively with 4 layers of both sides that rides over groove of folded filter paper of suitable dimension, as salt bridge.The film bar point sample of point sample is faced down, and the point sample place rides on the salt bridge towards cathode terminal, and the film bar needs to be adjacent to the exhibition of stretching tight with salt bridge.Energized, regulation voltage 100～260v, after 10～30 minutes, powered-down.Carry out isodynamic enzyme colour developing then: electrophoresis is crossed the film bar take out, the point sample face is attached on the above-mentioned colour developing offset plate, 37 ℃ of incubations 20～30 minutes, and promptly visible isozymogram.
The result: ALP4 can occur in the gestation, ALP4 appears in other people, and prompting has the tumour illness, and after showing with mentioned reagent, ALP4 disappears, and ALP5 occurs during liver disease, adds above-mentioned inhibitor ALP5 and disappears.Can differentiate internal organs source or the pathology location of ALP4, ALP5 thus.
Embodiment 6, do agarose gel electrophoresis, it includes: (1), compound concentration are 0.05mol/L barbitol buffer solution electrode solution, and pH is 8.6; (2), with glacial acetic acid as fixing agent, get 2.0ml packing ampoule E; (3), the preparation of Ago-Gel: it is the fine film damping fluid of 0.08mol/L bubble vinegar 100ml that agarose 0.7g is added concentration, heating for dissolving, packing while hot is prepared as ampoule A; (4) preparation colour developing liquid: colour developing liquid can be prepared as freeze-dried powder reagent, and it is with 0.5g substrate and 100ml damping fluid dissolving back packing ampoule B, freeze drying; With 0.7g diazol colours and 100ml damping fluid, dissolving back packing ampoule C, freeze drying, the time spent will be dissolved in water in the ampoule B, is dissolved in water in the ampoule C, with ampoule B, ampoule C contents mixed, is colour developing liquid.
Use: with ampoule A water proof heating for dissolving, water while hot on a glass plate, the filter paper of 0.3 * 1cm is put in the cooling back at the about 2cm of distance film one end place.Take off filter paper after about 10 minutes,, placed 5 minutes at these place's application of sample 8 microlitres.Then gel slab being placed on the electrophoresis truss that has added electrode solution, puts up a bridge with 4 layers of gauze in two ends, voltage 100v, electrophoresis 30 minutes, powered-down.Offset plate take out is added the above-mentioned colour developing liquid that has mixed, be placed in 37 ℃ of incubators incubation 20-30 minute, promptly visible isodynamic enzyme colour developing band.
The result: serum alkaline phosphatase (ALP) isodynamic enzyme can occur ALP1-6 bar district band from positive pole to negative pole, and ALP2 (liver type) and two kinds of isodynamic enzymes of ALP3 (bone type) appear in normal human serum.When primary carcinoma of liver, cirrhosis, liver type activity of isoenzyme strengthens.Bone type activity of isoenzyme strengthens when osteogenic sarcoma, hyperthyroidism.In the gestation, ALP4 can appear in serum, sharply descends if ALP4 is active, usually is the dead diagnostic markers of placenta insufficiency or fetus, and other people ALP4 occurs, are shown with the tumour illness in advance, and ALP1 and ALP6 occur when pathologic condition.
Claims (8)
1, cellulose acetate electrophoresis is done in a kind of preparation of alkaline phosphatase isoenzyme diagnostic kit, and it includes: (1), preparation 0.02～0.1mol/L electrode solution, and pH is 7.5-9.0; (2), with glacial acetic acid as fixing agent; (3), the fine film damping fluid of preparation 0.05～0.2mol/L bubble vinegar; It is characterized in that: in the fine film damping fluid of (3) described bubble vinegar packing ampoule A; With (2) described fixing agent packing ampoule E; (4) preparation colour developing liquid:
(a), compound concentration is 0.05～0.3mol/L damping fluid, pH is 9.0～11.0, among the packing ampoule B;
(b) with substrate and diazol colours packing ampoule C;
(c), with agarose or agar packing ampoule D; Content agarose or the agar among content substrate among the ampoule C and diazol colours, the ampoule D wherein, by weight percentage, substrate, diazol colours, agarose or agar are respectively 20.0%～86.7%, 10.5%～75.2%, 1.2%～28.6%, with concentration is that the damping fluid of 0.05～0.3mol/L is 100ml～250ml packing ampoule B, time spent with ampoule B add in the ampoule C, the content of ampoule D adds the water heating for dissolving, with the mixed liquor of ampoule B, ampoule C, add the interior mixing of ampoule D and be colour developing liquid then.
2, agarose gel electrophoresis is done in a kind of preparation of alkaline phosphatase isoenzyme diagnostic kit, and it includes:
(1), compound concentration is 0.02～0.1mol/L electrode solution, pH is 7.5-9.0;
(2), use glacial acetic acid as fixing agent; It is characterized in that: with (2) described fixing agent packing ampoule E;
(3), the preparation of Ago-Gel: it is the fine film damping fluid of 0.05～0.2mol/L bubble vinegar 80ml～120ml that agarose (or agar) 0.5g-1g is added concentration, heating for dissolving, packing while hot is prepared as ampoule A;
(4) preparation colour developing liquid
(a), compound concentration is 0.05～0.3mol/L damping fluid, pH is 9.0～11.0, among the packing ampoule B,
(b) with substrate and diazol colours packing ampoule C;
(c), with agarose or agar packing ampoule D; Content agarose or the agar among content substrate among the ampoule C and diazol colours, the ampoule D wherein, by weight percentage, substrate, diazol colours, agarose or agar are respectively 20.0%～86.7%, 10.5%～75.2%, 1.2%～28.6%, with concentration is that the damping fluid of 0.05～0.3mol/L is 100ml～250ml packing ampoule B, and the contents mixed of time spent with ampoule B, ampoule C, ampoule D is colour developing liquid.
3, the preparation of a kind of alkaline phosphatase isoenzyme diagnostic kit according to claim 1 and 2, in described colour developing liquid, add specific inhibitor packing ampoule F when it is characterized in that developing the color, wherein can add concentration among the ampoule F is 5～10mM L-phenylalanine, suppresses ALP4, ALP5; Or concentration is 5～10mM L-homoarginine, suppresses ALP2; Or concentration is 5～10mM L-tryptophane or leucine, suppresses ALP4; Or concentration is 3～5M urea, suppresses ALP3; Or concentration is the 1M ethylenediamine tetraacetic acid, suppresses ALP2, ALP3, ALP5.
4, the preparation of a kind of alkaline phosphatase isoenzyme diagnostic kit according to claim 1 and 2, it is characterized in that the damping fluid prepared among the described ampoule B, the damping fluid of the damping fluid of available Tris series, phosphoric acid series or borate buffer, diamido dimethyl 1.3 propylene glycol-HCL damping fluid.
5, the preparation of a kind of alkaline phosphatase isoenzyme diagnostic kit according to claim 1 and 2, it is characterized in that among the described ampoule C that substrate can be with the phosphate of naphthols or its derivant, as alpha-Naphthol phosphoric acid or betanaphthol phosphoric acid or alpha-Naphthol phosphoric acid salt compound, naphthols AS-MX phosphoric acid, naphthols AS-BI phosphoric acid, or its naphthols AS-MX phosphoric acid salt and naphthols AS-BI phosphoric acid salt, toluidine-5-bromine 3-indoles phosphoric acid, nitrobenzene-5-bromo-3-indoles phosphoric acid, tricresyl phosphate amine salt; Diazol colours can with solid blue B or solid blue BB, solid blue R or blue RR, solid purple B, three-bristle cudrania garnet GBC, fast red RC, fast red TR, six azos be paid magenta etc. admittedly.
6, the preparation of a kind of alkaline phosphatase isoenzyme diagnostic kit according to claim 1 and 2 is characterized in that the preparation of described colour developing liquid, dress damping fluid and substrate in the wherein said ampoule B, dissolving postlyophilization; It is that the 0.3g-1.5g substrate is added 100ml-250ml damping fluid dissolving back packing ampoule B.
7, the preparation of a kind of alkaline phosphatase isoenzyme diagnostic kit according to claim 1 and 2, it is characterized in that the preparation of described colour developing liquid, dress damping fluid and diazol colours in the wherein said ampoule C, the dissolving postlyophilization, it is the damping fluid dissolving back packing ampoule C that the diazol colours of 0.2g-1g is added 100ml-250ml.
8, the preparation of a kind of alkaline phosphatase isoenzyme diagnostic kit according to claim 2, the preparation that it is characterized in that described Ago-Gel also can be with agarose (or agar), sucrose and polyvinylpyrrolidone, be respectively 7.0%～32.3%, 55.6%～91.0%, 1.4%～19.4% by weight percentage, adding 100ml～250ml concentration is the fine film damping fluid of bubble vinegar of 0.05mol/L～0.2mol/L, heating for dissolving, packing while hot is prepared as ampoule A.
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| CN 01110756 CN1381725A (en) | 2001-04-20 | 2001-04-20 | Process for preparing diagnosing reagent kit of alkaline phosphatase and isozyme |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8476050B2 (en) | 2010-06-04 | 2013-07-02 | Shenzhen Mindray Bio-Medical Electronics Co., Ltd. | Conjugate preparation methods and related kit |
| CN101781675B (en) * | 2010-01-27 | 2013-08-14 | 福建泰普生物科学有限公司 | Target cell staining kit for displaying proliferative activity of cell and use method thereof |
| US8753858B2 (en) | 2009-12-31 | 2014-06-17 | Shenzhen Mindray Bio-Medical Electronics Co., Ltd. | Reagents and processes for stabilizing alkaline phosphatase or conjugates thereof |
| US8940512B2 (en) | 2010-06-04 | 2015-01-27 | Shenzhen Mindray Bio-Medical Electronics Co., Ltd. | Processes for synthesizing alkaline phosphatase conjugates |
-
2001
- 2001-04-20 CN CN 01110756 patent/CN1381725A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8753858B2 (en) | 2009-12-31 | 2014-06-17 | Shenzhen Mindray Bio-Medical Electronics Co., Ltd. | Reagents and processes for stabilizing alkaline phosphatase or conjugates thereof |
| CN101781675B (en) * | 2010-01-27 | 2013-08-14 | 福建泰普生物科学有限公司 | Target cell staining kit for displaying proliferative activity of cell and use method thereof |
| US8476050B2 (en) | 2010-06-04 | 2013-07-02 | Shenzhen Mindray Bio-Medical Electronics Co., Ltd. | Conjugate preparation methods and related kit |
| US8940512B2 (en) | 2010-06-04 | 2015-01-27 | Shenzhen Mindray Bio-Medical Electronics Co., Ltd. | Processes for synthesizing alkaline phosphatase conjugates |
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