CN101173909B - Serum protein electrophoresis gelose gel plate and production method thereof - Google Patents
Serum protein electrophoresis gelose gel plate and production method thereof Download PDFInfo
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- CN101173909B CN101173909B CN2006101141271A CN200610114127A CN101173909B CN 101173909 B CN101173909 B CN 101173909B CN 2006101141271 A CN2006101141271 A CN 2006101141271A CN 200610114127 A CN200610114127 A CN 200610114127A CN 101173909 B CN101173909 B CN 101173909B
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- agarose gel
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- agarose
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- plate
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Abstract
The invention relates to a gel slab of electrophoretic agarose of serum protein and the making method thereof. The gel slab is divided into two layers, the underlayer is a hydrophilic plastic slab, and the upper layer is a layer of agarose gel. The agarose gel is composed of agarose, barbital, barbital sodium, calcium lactate, trishydroxymethylaminomethane, ethylenediaminetetraacetic diasodium and boric acid. The making method is that: the agarose gel is prepared and put in the vessel, and heated in the water bath, after the agarose gel is completely dissolved, the temperature is kept constant, then the positioner is arranged on the heating plate with constant temperature, and enough agarose gel is sucked out by a pipette and squeezed under pressure into the mould, the mould fully poured by the agarose gel is arranged on the step cooling constant temperature board to be cooled and condensed to a semi-dry type status slowly, then the mould is opened, and the well paved gel slab is sealed well in the box. As the best matching reagent of the dry type plate electrophoresis instrument, the invention has the advantages of not only good consistency, but also convenient use, assembly and disassembly at any time, time-saving, clear and exact electrophoresis separation zone, good resolution, high precision and long service life.
Description
Technical field
The present invention relates to a kind of serum protein electrophoresis gelose gel plate and preparation method thereof, it is an employed carrier when doing haemocyanin dry type electrophoresis, belongs to external diagnosis reagent.
Background technology
Traditional manual electrophoresis method is with damping fluid in electrophoresis tank, make carrier with cellulose acetate membrane, by hand serum is added on the carrier, energized then, the beginning electrophoresis, its shortcoming is: it is pretty troublesome 1. to operate, and does the electrophoresis sample one time, needs a people just can finish in busy seven, eight hours; 2. electrophoresis is of low quality, and resolution is low, and the district is with not obvious and crooked, and low precision again can not long preservation; 3. data processing bothers, and electrophoresis is over and with scissors every district's band is cut off, and carries out wash-out separately in a kind of special solution, on tintmeter, carry out colorimetric washing the liquid that comes, be converted into number percent, this method is very inaccurate, and the trouble especially of calculate also; 4. uncontrollable its point sample amount of manual point sample, there is inconsistency in point sample, and the Zone electophoresis band is not on a line, and district's band that electrophoresis comes out is crooked.Electrophoretic separation technique can provide important evidence for clinical diagnosis in fact, but because method is backward, complicated operation, of low quality, the data that provide are inaccurate, make hospital not carry out electrophoretic separation technique gradually, hindered this subject development.
Summary of the invention
The object of the present invention is to provide a kind of easy to usely, save time, separating area belt is clear accurately, resolution height, long shelf-life, agarose gel plate of using as carrier when can be used for haemocyanin dry type electrophoretic separation and preparation method thereof.
The objective of the invention is to be realized that by following technical scheme agarose gel plate is a double-layer structure, bottom is the water wettability plastic plate, and the upper strata is an agarose gel, and is with no chemical reaction transparent adhesive tape bonding between two-layer.
The prescription of agarose gel is as follows:
Barbital (C
8H
12N
2O
3) 1～4 gram
Barbital sodium (C
8H
11N
2NaO
3) 10～15 grams
Calcium lactate (C
6H
10CaO
65H
2O) 0.3～1.5 gram
Trishydroxymethylaminomethane (C
4H
11NO
3) 10～14 grams
Disodium ethylene diamine tetraacetate (C
10H
14N
2Na
2O
82H
2O) 1.9～3 grams
Boric acid (H
3BO
3) 1.5～2.5 grams
Above-mentioned six kinds of components are dissolved in 1000 ml distilled waters by proportioning be mixed with solution
Contain 0.5～2.0 gram in the per 100 milliliters of above-mentioned solution of agarose
Its method for making may further comprise the steps:
1. earlier barbital, barbital sodium, calcium lactate, trishydroxymethylaminomethane, disodium ethylene diamine tetraacetate, boric acid are dissolved in the distilled water by said ratio, stir, fully dissolving is mixed with solution; Put into agarose by above-mentioned corresponding mixture ratio again and stir, will hold the container of above-mentioned mixed solution, be placed on heating in the water-bath, moisture-heat preservation treats that agarose all remains on 70 ℃ of-85 ℃ of constant temperature after the dissolving;
2. no chemical reaction transparent adhesive tape is coated in water wettability plastic plate upper face center position, the frame that stays 3 millimeter all around is gluing not, places then on the end template of mold, cope match-plate pattern is buckled again, and fixes with register pin.
3. the mold that fixes is placed on the heated at constant temperature plate, the temperature perseverance is at 50-65 ℃;
4. fall glue, the agarose gel with special-purpose aspirator sucking-off q.s adds extrusion in mold, and under the effect of pressure, agarose gel can the interior gas of smooth and easy emptying mold and be full of the mold space;
5. the mold that waters full agarose gel is placed on the ladder cooling temperature-constant plate, steady cooling slowly makes the agarose gel half dry type state that slowly congeals into;
6. with specific purpose tool mold is opened after 20 minutes, the gel slab of completing is tested, excellent can being placed in the box sealed, and seals with aluminium plastic bag again.
Described method for making step 4. in used pressure, just use the hand squeezing pressure.
Described gel slab thickness is at 0.3～0.9 millimeter, and evenly uneven phenomenon can not occur, and is can not became uneven even, if above-mentioned phenomenon just can't guarantee the quality of Zone electophoresis band.So it is very strict to profile pattern, uniformity requirement.Gel slab is generally square, also can be rectangle, and the top-layer agar carbohydrate gum is than all little 3 millimeter around the bottom water wettability plastic plate.
The transparent adhesive tape of described no chemical reaction can be gelatin or pig skin gelatin etc.
Used mold is dismantled and assembled, and when the water wettability plastic plate placed the mold end template, the water wettability plastic plate had been coated with the transparent adhesive tape that has or not chemical reaction, and therefore, when agarose gel took the mold space, its bottom promptly was bonded into one with the water wettability plastic plate.
The reasonable science that the molding fetal venthole will be opened does not produce the dead angle, and liquid agarose gel can well-proportionedly be discharged gas, thereby makes the yet just very even of agarose gel thin layer stream.
Inventive principle: contain 100 multiple proteins in the human blood, each component concentration has different separately functions, and various specific variable quantity can occur under different pathological conditions, thereby judges the different causes of disease.Since the foundation of Tiselies interface movement electrophoresis and the use of zone electrophoresis appears subsequently and the appearance of autophoresis instrument since, the protein electrophoresis technology obtains greatly progress, serum protein electrophoresis has important value at laboratory medicine.
Serum proteins belong to ampholyte, and when the pH value of solution protein belt negative charge during greater than isoelectric points of proteins, under electric field action, anode moves during electrophoresis.With agarose gel plate as carrier, because protein institute is with the difference of net charge quantity isoelectric point and the molecular weight different mobility of generation when the electrophoresis, make haemocyanin isolate Wu Tiao district band, be albumin, alpha1 Globulin, alpha2 Globulin, beta Globulin and gamma Globulin, the relative ratio of these protein components provides valuable clinical foundation for the diagnosis and the prognosis of some disease.
The present invention must be in good Zone electophoresis strap of could swimming out in the half dry type state under electric field action, district's band that the sharpness of swimming out is high so must pack, just can be opened during use.
The present invention is placed on the electrophoresis flat board of dry type disk electrophoresis instrument to use.
Advantage of the present invention is:
1, surfacing, thickness is even, and thickness is only at 0.3～0.9 millimeter.
2, it is the matched reagent of dry type disk electrophoresis instrument the best, long service life not only, and high conformity, and also easy to use, promptly tear i.e. usefulness open, do not need oneself to make, to save time, separating area belt is clear accurately, the resolution height.
3, the data that provide are accurate, can provide important evidence for clinical diagnosis.
Description of drawings
Fig. 1 is the serum protein electrophoresis gelose gel plate planar structure synoptic diagram of bowing
Fig. 2 is the A-A sectional view of Fig. 1
Fig. 3 is the positioner structure synoptic diagram
Among the figure: 1. water wettability plastic plate, 2. agarose gel, 3. end template, 4. cope match-plate pattern, 5. register pin, 6. plastic squeeze hole, 7. venthole; Arrow is respectively plastic squeeze direction and outgassing direction.
Embodiment
Embodiment 1
The described agarose gel plate of present embodiment is a double-layer structure, and bottom is a water wettability plastic plate 1, and the upper strata is an agarose gel 2, two-layer between with no chemical reaction transparent adhesive tape bonding.
The prescription of agarose gel 2 is as follows:
Barbital (C
8H
12N
2O
3) 1 gram
Barbital sodium (C
8H
11N
2NaO
3) 12 grams
Calcium lactate (C
6H
10CaO
65H
2O) 1.5 grams
Trishydroxymethylaminomethane (C
4H
11NO
3) 10 grams
Disodium ethylene diamine tetraacetate (C
10H
14N
2Na
2O
82H
2O) 2.4 grams
Boric acid (H
3BO
3) 2.5 grams
Above-mentioned six kinds of components are dissolved in 1000 ml distilled waters by proportioning be mixed with solution
Contain 0.5 gram in the per 100 milliliters of above-mentioned solution of agarose
Its method for making may further comprise the steps:
1. earlier barbital, barbital sodium, calcium lactate, trishydroxymethylaminomethane, disodium ethylene diamine tetraacetate, boric acid are dissolved in the distilled water by said ratio, stir, fully dissolving is mixed with solution; Put into agarose by above-mentioned corresponding mixture ratio again and stir, will hold the container of above-mentioned mixed solution, be placed on heating in the water-bath, moisture-heat preservation treats that agarose all remains on 70 ℃ of-85 ℃ of constant temperature after the dissolving;
2. no chemical reaction transparent adhesive tape is coated in water wettability plastic plate upper face center position, the frame that stays 3 millimeter all around is gluing not, places then on the end template of mold, cope match-plate pattern is buckled again, and fixes with register pin.
3. the mold that fixes is placed on the heated at constant temperature plate, the temperature perseverance is at 50-65 ℃;
4. fall glue, the agarose gel with special-purpose aspirator sucking-off q.s adds extrusion in mold, and under the effect of pressure, agarose gel can the interior gas of smooth and easy emptying mold and be full of the mold space;
5. the mold that waters full agarose gel is placed on the ladder cooling temperature-constant plate, steady cooling slowly makes the agarose gel half dry type state that slowly congeals into;
6. with specific purpose tool mold is opened after 20 minutes, the gel slab of completing is tested, excellent can being placed in the box sealed; Seal with aluminium plastic bag again.
Described gel slab is a square, and thickness is at 0.3～0.9 millimeter, and the uneven phenomenon of the even no concave-convex of thickness; Top-layer agar carbohydrate gum 2 is than all little 3 millimeters around the bottom water wettability plastic plate 1.
Embodiment 2:
The described agarose gel plate of present embodiment is a double-layer structure, and bottom is a water wettability plastic plate 1, and the upper strata is an agarose gel 2, two-layer between with no chemical reaction transparent adhesive tape bonding.
The prescription of agarose gel 2 is as follows:
Barbital (C
8H
12N
2O
3) 3 grams
Barbital sodium (C
8H
11N
2NaO
3) 15 grams
Calcium lactate (C
6H
10CaO
65H
2O) 0.3 gram
Trishydroxymethylaminomethane (C
4H
11NO
3) 12 grams
Disodium ethylene diamine tetraacetate (C
10H
14N
2Na
2O
82H
2O) 3 grams
Boric acid (H
3BO
3) 1.5 grams
Above-mentioned six kinds of components are dissolved in 1000 ml distilled waters by proportioning be mixed with solution
Contain 1.2 grams in the per 100 milliliters of above-mentioned solution of agarose
Its method for making is with embodiment 1.
Embodiment 3:
The described agarose gel plate of present embodiment is a double-layer structure, and bottom is a water wettability plastic plate 1, and the upper strata is an agarose gel 2, two-layer between with no chemical reaction transparent adhesive tape bonding.
The prescription of agarose gel 2 is as follows:
Barbital (C
8H
12N
2O
3) 4 grams
Barbital sodium (C
8H
11N
2NaO
3) 10 grams
Calcium lactate (C
6H
10CaO
65H
2O) 0.9 gram
Trishydroxymethylaminomethane (C
4H
11NO
3) 14 grams
Disodium ethylene diamine tetraacetate (C
10H
14N
2Na
2O
82H
2O) 1.9 grams
Boric acid (H
3BO
3) 2 grams
Above-mentioned six kinds of components are dissolved in 1000 ml distilled waters by proportioning be mixed with solution
Contain 2.0 grams in the per 100 milliliters of above-mentioned solution of agarose
Its method for making is with embodiment 1.
Claims (2)
1. serum protein electrophoresis gelose gel plate, it is characterized in that: gel slab is a double-layer structure, and bottom is water wettability plastic plate (1), and the upper strata is agarose gel (2), and is bonding with no chemical reaction transparent adhesive tape between two-layer;
Described gel slab thickness is at 0.3～0.9 millimeter;
The prescription of described agarose gel (2) is as follows:
Barbital 1～4 gram
Barbital sodium 10～15 grams
Calcium lactate 0.3～1.5 gram
Trishydroxymethylaminomethane 10～14 grams
Disodium ethylene diamine tetraacetate 1.9～3 grams
Boric acid 1.5～2.5 grams
Above-mentioned six kinds of components are dissolved in 1000 ml distilled waters by proportioning be mixed with solution;
Contain 0.5～2.0 gram in the per 100 milliliters of above-mentioned solution of agarose.
2. the method for making of a serum protein electrophoresis gelose gel plate is characterized in that: may further comprise the steps:
1. earlier barbital, barbital sodium, calcium lactate, trishydroxymethylaminomethane, disodium ethylene diamine tetraacetate, boric acid are dissolved in the distilled water by the described proportioning of claim 1, stir, fully dissolving is mixed with solution; Put into agarose by the described corresponding mixture ratio of claim 1 again and stir, will hold the container of above-mentioned mixed solution, be placed on heating in the water-bath, moisture-heat preservation treats that agarose all remains on 70 ℃ of-85 ℃ of constant temperature after the dissolving;
2. will coat with water wettability plastic plate and agarose gel in water wettability plastic plate upper face center position does not have the chemical reaction transparent adhesive tape, and the frame that stays 3 millimeter all around is gluing not, places then on the end template of mold, cope match-plate pattern is buckled again, and fixes with register pin;
3. the mold that fixes is placed on the heated at constant temperature plate, the temperature perseverance is at 50-65 ℃;
4. fall glue, the agarose gel with special-purpose aspirator sucking-off q.s adds extrusion in mold, and under the effect of pressure, agarose gel can the interior gas of smooth and easy emptying mold and be full of the mold space;
5. the mold that waters full agarose gel is placed on the ladder cooling temperature-constant plate, steady cooling slowly makes the agarose gel half dry type state that slowly congeals into;
6. with specific purpose tool mold is opened after 20 minutes, the gel slab of completing is tested, excellent promptly being placed in the box sealed; Seal with aluminium plastic bag again.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2006101141271A CN101173909B (en) | 2006-10-30 | 2006-10-30 | Serum protein electrophoresis gelose gel plate and production method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2006101141271A CN101173909B (en) | 2006-10-30 | 2006-10-30 | Serum protein electrophoresis gelose gel plate and production method thereof |
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| Publication Number | Publication Date |
|---|---|
| CN101173909A CN101173909A (en) | 2008-05-07 |
| CN101173909B true CN101173909B (en) | 2011-01-05 |
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|---|---|---|---|
| CN2006101141271A Expired - Fee Related CN101173909B (en) | 2006-10-30 | 2006-10-30 | Serum protein electrophoresis gelose gel plate and production method thereof |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104597098A (en) * | 2013-12-20 | 2015-05-06 | 上海迪安医学检验所有限公司 | Agarose gel plate storage method |
| CN104597099A (en) * | 2013-12-20 | 2015-05-06 | 上海迪安医学检验所有限公司 | Antiseptic agarose gel plate preparation method |
| CN105699647B (en) * | 2015-09-11 | 2018-10-12 | 中国农业科学院哈尔滨兽医研究所 | A kind of novel electrophoretic buffer and its preparation method and application |
| CN111777659B (en) * | 2020-07-09 | 2023-11-24 | 向开军 | Universal electric field driven protein solid phase renaturation method |
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|---|---|---|---|---|
| US6569306B1 (en) * | 2000-04-10 | 2003-05-27 | Amersham Pharmacia Biotech, Inc. | Cassette for gel electrophoresis having solid buffer reservoirs |
| CN1431315A (en) * | 2003-01-09 | 2003-07-23 | 中国人民解放军第二军医大学 | Kit for detecting lactate dehydrogenase isoenzyme |
| CN2601403Y (en) * | 2003-02-21 | 2004-01-28 | 郭晏海 | Agarose gel plate forming appts. |
-
2006
- 2006-10-30 CN CN2006101141271A patent/CN101173909B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6569306B1 (en) * | 2000-04-10 | 2003-05-27 | Amersham Pharmacia Biotech, Inc. | Cassette for gel electrophoresis having solid buffer reservoirs |
| CN1431315A (en) * | 2003-01-09 | 2003-07-23 | 中国人民解放军第二军医大学 | Kit for detecting lactate dehydrogenase isoenzyme |
| CN2601403Y (en) * | 2003-02-21 | 2004-01-28 | 郭晏海 | Agarose gel plate forming appts. |
Non-Patent Citations (6)
| Title |
|---|
| JP昭58-103656A 1983.06.20 |
| JP特开2001-2706A 2001.01.09 |
| 宋宏伟等.石英芯片电泳分离检测尿中蛋白的研究.分析测试学报25 4.2006,25(4),33. |
| 宋宏伟等.石英芯片电泳分离检测尿中蛋白的研究.分析测试学报25 4.2006,25(4),33. * |
| 赵绍林等.超薄琼脂糖凝胶的简易制备技术.临床检验杂志22 4.2004,22(4),251. |
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|---|---|
| CN101173909A (en) | 2008-05-07 |
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Granted publication date: 20110105 Termination date: 20181030 |