CN105987839B - A kind of EDTA- citric acid antigens reparation liquid - Google Patents
A kind of EDTA- citric acid antigens reparation liquid Download PDFInfo
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- CN105987839B CN105987839B CN201610385941.0A CN201610385941A CN105987839B CN 105987839 B CN105987839 B CN 105987839B CN 201610385941 A CN201610385941 A CN 201610385941A CN 105987839 B CN105987839 B CN 105987839B
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- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Natural products OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 title claims abstract description 90
- 239000000427 antigen Substances 0.000 title claims abstract description 73
- 102000036639 antigens Human genes 0.000 title claims abstract description 73
- 108091007433 antigens Proteins 0.000 title claims abstract description 73
- 239000007788 liquid Substances 0.000 title claims abstract description 57
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims abstract description 47
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 43
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims abstract description 38
- LTUDISCZKZHRMJ-UHFFFAOYSA-N potassium;hydrate Chemical compound O.[K] LTUDISCZKZHRMJ-UHFFFAOYSA-N 0.000 claims abstract description 34
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims abstract description 33
- 239000007983 Tris buffer Substances 0.000 claims abstract description 31
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims abstract description 31
- 230000008439 repair process Effects 0.000 claims abstract description 30
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 22
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 claims abstract description 19
- 241001474374 Blennius Species 0.000 claims description 47
- 241000195493 Cryptophyta Species 0.000 claims description 27
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 26
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 26
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 26
- 239000000872 buffer Substances 0.000 claims description 19
- 238000002360 preparation method Methods 0.000 claims description 13
- 239000002002 slurry Substances 0.000 claims description 13
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 12
- 238000000605 extraction Methods 0.000 claims description 12
- 230000001954 sterilising effect Effects 0.000 claims description 12
- 239000011268 mixed slurry Substances 0.000 claims description 11
- 239000000243 solution Substances 0.000 claims description 10
- 230000005855 radiation Effects 0.000 claims description 8
- 235000005979 Citrus limon Nutrition 0.000 claims description 7
- 238000004140 cleaning Methods 0.000 claims description 7
- 238000004090 dissolution Methods 0.000 claims description 7
- 244000131522 Citrus pyriformis Species 0.000 claims description 6
- 235000019750 Crude protein Nutrition 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 239000002253 acid Substances 0.000 claims description 4
- YASYEJJMZJALEJ-UHFFFAOYSA-N Citric acid monohydrate Chemical class O.OC(=O)CC(O)(C(O)=O)CC(O)=O YASYEJJMZJALEJ-UHFFFAOYSA-N 0.000 claims description 2
- 229960004106 citric acid Drugs 0.000 claims 7
- OAGSFHDUINSAMQ-UHFFFAOYSA-N 2-hydroxypropane-1,2,3-tricarboxylic acid;sodium;hydrate Chemical compound O.[Na].OC(=O)CC(O)(C(O)=O)CC(O)=O OAGSFHDUINSAMQ-UHFFFAOYSA-N 0.000 claims 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims 1
- 229910052708 sodium Inorganic materials 0.000 claims 1
- 239000011734 sodium Substances 0.000 claims 1
- 238000003364 immunohistochemistry Methods 0.000 abstract description 11
- 230000000694 effects Effects 0.000 abstract description 10
- 238000004043 dyeing Methods 0.000 abstract description 7
- 238000004040 coloring Methods 0.000 abstract description 3
- 230000008595 infiltration Effects 0.000 abstract description 3
- 238000001764 infiltration Methods 0.000 abstract description 3
- 235000019441 ethanol Nutrition 0.000 description 17
- 238000000034 method Methods 0.000 description 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 8
- 238000010025 steaming Methods 0.000 description 5
- -1 ethyl alcohol Amine Chemical class 0.000 description 4
- 235000011187 glycerol Nutrition 0.000 description 4
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 230000008901 benefit Effects 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- JAHNSTQSQJOJLO-UHFFFAOYSA-N 2-(3-fluorophenyl)-1h-imidazole Chemical compound FC1=CC=CC(C=2NC=CN=2)=C1 JAHNSTQSQJOJLO-UHFFFAOYSA-N 0.000 description 1
- OFNISBHGPNMTMS-UHFFFAOYSA-N 3-methylideneoxolane-2,5-dione Chemical compound C=C1CC(=O)OC1=O OFNISBHGPNMTMS-UHFFFAOYSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 244000248349 Citrus limon Species 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- HNEGQIOMVPPMNR-IHWYPQMZSA-N citraconic acid Chemical compound OC(=O)C(/C)=C\C(O)=O HNEGQIOMVPPMNR-IHWYPQMZSA-N 0.000 description 1
- 229940018557 citraconic acid Drugs 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 230000000093 cytochemical effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 238000005470 impregnation Methods 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- LVHBHZANLOWSRM-UHFFFAOYSA-N methylenebutanedioic acid Natural products OC(=O)CC(=C)C(O)=O LVHBHZANLOWSRM-UHFFFAOYSA-N 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 238000010827 pathological analysis Methods 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
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- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- General Physics & Mathematics (AREA)
- Urology & Nephrology (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
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Abstract
The present invention relates to immunohistochemistry technology fields, disclose a kind of EDTA citric acid antigens reparation liquid, and the EDTA citric acid antigens per 1000mL repair liquid and include：Tris Base 1.20 1.25g, EDTA 0.35 0.40g, 0.35 0.40g of monohydrate potassium, 2.40 2.42g of trisodium citrate dihydrate, 400 600 microlitres of polysorbas20, triethanolamine or ethylene glycol or 50 500mL of trihydroxylic alcohol and the single of surplus steam water.The EDTA citric acid antigens reparation liquid infiltration of the present invention, repairing effect are good, can be repaired to histotomy under less dosage, histotomy is easy dyeing after reparation, and coloring is good.And it is not volatile to repair liquid, and full-automatic immunohistochemistry instrument can be coordinated to use.
Description
Technical field
The present invention relates to immunohistochemistry technology fields more particularly to a kind of EDTA- citric acid antigens to repair liquid.
Background technology
Immunohistochemistry technology is the detection technique that pathological diagnosis in recent years rapidly develops, as a kind of important pathology
Aided diagnosis method is paid more and more attention in clinical application.It is right with the application of immunohistochemistry autostainer
It is also increasingly improved in the requirement of dyeing quality.Wherein, antigen retrieval has of crucial importance during entire immunohistochemical staining
Effect, the quality of repairing condition directly decides the quality of final coloration result.Full-automatic dyeing instrument is because of the knot of its own
Structure feature, the container volume for being used to hold reparation liquid is smaller, histotomy cannot be fully immersed in as repaired by hand
The condition in liquid is repaired, and immunohistochemistry autostainer is needed in antigen retrieval in higher temperature（100 DEG C of left sides
It is right）Lower progress, therefore reparation liquid is readily volatilized, further reduces the volume for repairing liquid.Thus, how in full-automatic immune group
It is the problem that immunohistochemistry autostainer must solve to complete normal antigen retrieval on change dyeing instrument.
But since the current immunohistochemistry autostainer country is not yet common, few people use, therefore nobody
Notice above-mentioned technical problem.The external method for solving above-mentioned technical problem is typically to control the environment of antigen retrieval at present, such as
Control atmospheric pressure, environment temperature etc., these solution complicated conditions, it is difficult to practical application.It does not find by changing also at present
The report of above-mentioned technical problem is solved into antigen retrieval buffers.
Current antigen retrieval buffers have a large amount in variety, as EDTA antigen retrieval buffers, citric acid antigen repair liquid and EDTA-
Citric acid compounds antigen retrieval buffers etc..Application No. is 201180024136.0 Chinese patents to disclose a kind of antigen retrieval buffers
With antigen retrieval method, the antigen retrieval being used in the cytochemical staining procedure of immuning tissue and cell.The antigen retrieval buffers are
Itaconic acid, itaconic anhydride or citraconic acid aqueous solution.The antigen retrieval method is using above-mentioned antigen retrieval buffers to histotomy
Carry out high temperature or high temperature and pressure reparation.The antigen retrieval buffers and antigen retrieval method of offer are high with dyeing quality, reproducibility,
The good advantage of safety, stability.
The solvent of above-mentioned antigen retrieval buffers is water, if applied on immunohistochemistry autostainer, on the one hand, this is anti-
Original repairs liquid and needs the repairing effect in the case where dosage is big better, can not can not be by histotomy thorough impregnation in less dosage
In the case of complete preferable repair；On the other hand, low boiling point, it is readily volatilized at relatively high temperatures, it is easy to cause tissue and cuts
Desiccation situation occurs for piece, and desiccation once occurs, and the antigen of histotomy is changing to irreversible destruction and can not repair.
Invention content
In order to solve the above technical problem, the present invention provides a kind of EDTA- citric acid antigens to repair liquid.The present invention's
The infiltration of EDTA- citric acid antigens reparation liquid, repairing effect are good, can repair, repair to histotomy under less dosage
Histotomy is easy dyeing after multiple, and coloring is good.And it is not volatile to repair liquid, and full-automatic immunohistochemistry instrument can be coordinated to make
With.
The specific technical solution of the present invention is：A kind of EDTA- citric acid antigens reparation liquid, the EDTA- lemons per 1000mL
Sour antigen retrieval buffers include：Tris-Base 1.20-1.25g, EDTA 0.35-0.40g, monohydrate potassium 0.35-
0.40g, trisodium citrate dihydrate 2.40-2.42g, 400-600 microlitres of Tween-20, triethanolamine or ethylene glycol or trihydroxylic alcohol
50-500mL and the single of surplus steam water.
In the antigen retrieval buffers of the present invention, repaiied using EDTA, monohydrate potassium, trisodium citrate dihydrate as main
Active ingredient, and water is steamed as mixed solvent with list using triethanolamine or ethylene glycol or trihydroxylic alcohol.Wherein, triethanolamine or
The addition of ethylene glycol or trihydroxylic alcohol can improve the boiling point of antigen retrieval buffers, prevent it from excessively volatilizing.And triethanolamine or second
Compared with the solvent of other high points, advantage is that the influence to the active constituent in antigen retrieval buffers is smaller for glycol or trihydroxylic alcohol,
And it is preferable with the compatibility of histotomy, toxicity is low, will not cause excessively to influence on the reparation of histotomy, wherein trihydroxylic alcohol,
The effect of triethylamine alcohol is better than ethylene glycol.Tween-20 plays emulsification, can improve the compatible of each substance in antigen retrieval buffers
Property.
Preferably, the EDTA- citric acid antigens reparation liquid per 1000mL includes：Tris-Base 1.2114g, EDTA
0.37224g, monohydrate potassium 0.378252g, trisodium citrate dihydrate 2.411784g, 500 microlitres of Tween-20, three ethyl alcohol
Amine or ethylene glycol or trihydroxylic alcohol 150mL and the single of surplus steam water.
Preferably, the EDTA- citric acid antigens reparation liquid per 1000mL includes：Tris-Base 1.2114g, EDTA
0.37224g, monohydrate potassium 0.378252g, trisodium citrate dihydrate 2.411784g, 500 microlitres of Tween-20, three ethyl alcohol
Amine 150mL and the single of surplus steam water.
Preferably, the EDTA- citric acid antigens reparation liquid per 1000mL includes：Tris-Base 1.20-1.25g,
EDTA 0.35-0.40g, monohydrate potassium 0.378252g, trisodium citrate dihydrate 2.411784g, Tween-20 400-600
Microlitre, single steaming water of triethanolamine or ethylene glycol or trihydroxylic alcohol 50-500mL, marine algae extract 0.5-1.5g and surplus.
Preferably, the EDTA- citric acid antigens reparation liquid per 1000mL includes：Tris-Base 1.2114g, EDTA
0.37224g, monohydrate potassium 0.378252g, trisodium citrate dihydrate 2.411784g, 500 microlitres of Tween-20, three ethyl alcohol
The single of amine or ethylene glycol or trihydroxylic alcohol 150mL, marine algae extract 1g and surplus steams water.
Preferably, the EDTA- citric acid antigens reparation liquid per 1000mL includes：Tris-Base 1.2114g, EDTA
0.37224g, monohydrate potassium 0.378252g, trisodium citrate dihydrate 2.411784g, 500 microlitres of Tween-20, three ethyl alcohol
Amine 150mL, marine algae extract 1g and the single of surplus steam water.
Preferably, the preparation method that the EDTA- citric acid antigens repair liquid is as follows：
Tris-Base, EDTA, monohydrate potassium, trisodium citrate dihydrate are weighed according to the ratio, steam water dissolution with single
After Tris-Base, EDTA, monohydrate potassium, trisodium citrate dihydrate, the triethanolamine or ethylene glycol or three of formula ratio is added
First alcohol adds Tween-20 after being completely dissolved, be finally settled to rated capacity with single water that steams.
Preferably, the preparation method that the EDTA- citric acid antigens repair liquid also can be as follows：Tris- is weighed according to the ratio
Base, EDTA, monohydrate potassium, trisodium citrate dihydrate steam water dissolution Tris-Base, EDTA, a citrate hydrate with single
Acid after trisodium citrate dihydrate, is added the triethanolamine or ethylene glycol or trihydroxylic alcohol of formula ratio, seaweed is added after being completely dissolved and is carried
Object and Tween-20 are taken, is finally settled to rated capacity with single water that steams.
Preferably, the preparation method of the marine algae extract is：By the seaweed after cleaning and water 2-4 in mass ratio:10
Mixing, is used in combination beater that seaweed slurry is made；By hydrochloric acid solution that seaweed slurry and pH value are 3-5 by volume 1:1-2 is mixed,
Mixed slurry is obtained, microwave radiation exaraction is carried out to mixed slurry, solid is filtered to remove after extraction and obtains seaweed extracted liquor, it is right
Seaweed extracted liquor cures sterilizing 0.5-1.5h in the environment of 100-110 DEG C, 4-8MPa, can make part more in maturing process
Sugar degradation；Seaweed extracted liquor after sterilizing to curing is filtered the crude protein that removal has been denaturalized；Filtered seaweed is extracted
Liquid pH is adjusted to neutrality, and marine algae extract is made after being finally freeze-dried.
In order to further optimize, marine algae extract is also added in antigen retrieval buffers of the invention.It is obtained in the present invention
Marine algae extract contains the substances such as natural alginate and natural polysaccharide, and biological mildness is good, with histotomy compatibility
It is good.A small amount of marine algae extract viscosity soluble in water that liquid can be made certain, water lock effect is good, can further prevent antigen
Solvent in liquid is repaired to volatilize.In addition, marine algae extract to the repairing activity ingredient in antigen retrieval buffers can physical bond, seaweed
After extract is contacted with histotomy, one layer of thin mucous membrane can be formed on histotomy surface, so that in connection
Repairing activity ingredient can be contacted fully with histotomy, in the case of less dosage antigen retrieval buffers, improve repairing effect.
But the used in amounts of marine algae extract wants stringent control, if dosage is excessive, antigen retrieval fluid viscosity can be caused excessively high, mobility
It is too poor, and combined excessively with histotomy, it is not easy subsequent cleaning.
Preferably, the condition of the microwave radiation exaraction is：70-90 DEG C of temperature, time 5-10min, microwave power
800-1000W, microwave frequency 2450MHZ.
Preferably, the trihydroxylic alcohol can be glycerine.
It is compared with the prior art, the beneficial effects of the invention are as follows：
The EDTA- citric acid antigens reparation liquid infiltration of the present invention, repairing effect are good, can be under less dosage to tissue
Slice is repaired.Reparation liquid needed for average every histotomy only needs 100-150uL.
Histotomy is easy dyeing after reparation, and coloring is good.
And it is not volatile to repair liquid, and full-automatic immunohistochemistry instrument can be coordinated to use.
Specific implementation mode
With reference to embodiment, the invention will be further described.
Monohydrate potassium 0.35-0.40g, trisodium citrate dihydrate 2.40-2.42g,
Monohydrate potassium 0.378252g, trisodium citrate dihydrate 2.411784g,
Embodiment 1
A kind of EDTA- citric acid antigens reparation liquid, the EDTA- citric acid antigens per 1000mL repair liquid and include：
Tris-Base 1.2114g, EDTA 0.37224g, monohydrate potassium 0.378252g, trisodium citrate dihydrate
The single of 2.411784g, 500 microlitres of Tween-20, triethanolamine 150mL and surplus steams water.
Preparation method is as follows：Tris-Base, EDTA, monohydrate potassium, trisodium citrate dihydrate are weighed according to the ratio, are used
After single steaming water dissolution Tris-Base, EDTA, monohydrate potassium, trisodium citrate dihydrate, the triethanolamine of formula ratio is added,
Tween-20 is added after being completely dissolved, and is finally settled to rated capacity with single water that steams.
Embodiment 2
A kind of EDTA- citric acid antigens reparation liquid, the EDTA- citric acid antigens per 1000mL repair liquid and include：
Tris-Base 1.2114g, EDTA 0.37224g, monohydrate potassium 0.378252g, trisodium citrate dihydrate
The single of 2.411784g, 500 microlitres of Tween-20, ethylene glycol 325mL and surplus steams water.
Preparation method is as follows：Tris-Base, EDTA, monohydrate potassium, trisodium citrate dihydrate are weighed according to the ratio, are used
After single steaming water dissolution Tris-Base, EDTA, monohydrate potassium, trisodium citrate dihydrate, the ethylene glycol of formula ratio is added, it is complete
Tween-20 is added after fully dissolved, is finally settled to rated capacity with single water that steams.
Embodiment 3
A kind of EDTA- citric acid antigens reparation liquid, the EDTA- citric acid antigens per 1000mL repair liquid and include：Tris-
Base 1.20g, EDTA 0.35g, monohydrate potassium 0.35g, trisodium citrate dihydrate 2.40g, 400 microlitres of Tween-20,
Triethanolamine 50mL and the single of surplus steam water.
Embodiment 4
A kind of EDTA- citric acid antigens reparation liquid, the EDTA- citric acid antigens per 1000mL repair liquid and include：Tris-
Base 1.25g, EDTA 0.40g, monohydrate potassium 0.40g, trisodium citrate dihydrate 2.42g, 600 microlitres of Tween-20,
Triethanolamine 500mL and the single of surplus steam water.
Embodiment 5
A kind of EDTA- citric acid antigens reparation liquid, the EDTA- citric acid antigens per 1000mL repair liquid and include：
Tris-Base 1.2114g, EDTA 0.37224g, monohydrate potassium 0.378252g, trisodium citrate dihydrate
The single of 2.411784g, 500 microlitres of Tween-20, glycerine 300mL and surplus steams water.
Embodiment 6
A kind of EDTA- citric acid antigens reparation liquid, the EDTA- citric acid antigens per 1000mL repair liquid and include：
Tris-Base 1.2114g, EDTA 0.37224g, monohydrate potassium 0.378252g, trisodium citrate dihydrate
The single of 2.411784g, 500 microlitres, triethanolamine 150mL, marine algae extract 1g of Tween-20 and surplus steams water.
Preparation method is as follows：Tris-Base, EDTA, monohydrate potassium, trisodium citrate dihydrate are weighed according to the ratio, are used
After single steaming water dissolution Tris-Base, EDTA, monohydrate potassium, trisodium citrate dihydrate, the triethanolamine of formula ratio is added,
Marine algae extract and Tween-20 are added after being completely dissolved, and are finally settled to rated capacity with single water that steams.
The preparation method of the marine algae extract is：By the seaweed after cleaning and water in mass ratio 3:10 mixing, are used in combination and beat
Seaweed slurry is made in pulp grinder；The hydrochloric acid solution by volume 1 for being 4 by seaweed slurry and pH value:1.5 mixing, obtain mixed slurry,
Microwave radiation exaraction is carried out to mixed slurry, wherein extraction conditions are：80 DEG C, time 7.5min of temperature, microwave power 900W are micro-
Wave frequency rate 2450MHZ.It is filtered to remove solid after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 105 DEG C, the ring of 6MPa
Curing sterilizing 1h under border；Seaweed extracted liquor after sterilizing to curing is filtered removal crude protein；Filtered seaweed is extracted
Liquid pH is adjusted to neutrality, and marine algae extract is made after being finally freeze-dried.
Embodiment 7
A kind of EDTA- citric acid antigens reparation liquid, the EDTA- citric acid antigens per 1000mL repair liquid and include：Often
The EDTA- citric acid antigens of 1000mL repair liquid：Tris-Base 1.2114g, EDTA 0.37224g, a hydration lemon
Lemon acid 0.378252g, trisodium citrate dihydrate 2.411784g, 500 microlitres, triethanolamine 150mL of Tween-20, seaweed extraction
Object 0.5g and the single of surplus steam water.
The preparation method of the marine algae extract is：By the seaweed after cleaning and water in mass ratio 2:10 mixing, are used in combination and beat
Seaweed slurry is made in pulp grinder；By hydrochloric acid solution that seaweed slurry and pH value are 3-5 by volume 1:1 mixing, obtains mixed slurry,
Microwave radiation exaraction is carried out to mixed slurry, wherein extraction conditions are：Temperature 70 C, time 10min, microwave power 800W are micro-
Wave frequency rate 2450MHZ.It is filtered to remove solid after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 100 DEG C, 8MPa's
Curing sterilizing 0.5h under environment；Seaweed extracted liquor after sterilizing to curing is filtered removal crude protein；By filtered seaweed
Extracting solution pH is adjusted to neutrality, and marine algae extract is made after being finally freeze-dried.
Embodiment 8
EDTA- citric acid antigens per 1000mL repair liquid：Tris-Base 1.2114g, EDTA
0.37224g, monohydrate potassium 0.378252g, trisodium citrate dihydrate 2.411784g, 500 microlitres of Tween-20, three ethyl alcohol
Amine 150mL, marine algae extract 1.5g and the single of surplus steam water.
The preparation method of the marine algae extract is：By the seaweed after cleaning and water in mass ratio 4:10 mixing, are used in combination and beat
Seaweed slurry is made in pulp grinder；By hydrochloric acid solution that seaweed slurry and pH value are 3-5 by volume 1:2 mixing, obtain mixing slurry
Material carries out microwave radiation exaraction to mixed slurry, and wherein extraction conditions are：90 DEG C, time 5min, microwave power 1000W of temperature,
Microwave frequency 2450MHZ.It is filtered to remove solid after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 110 DEG C, 4MPa's
Curing sterilizing 1.5h under environment；Seaweed extracted liquor after sterilizing to curing is filtered removal crude protein；By filtered seaweed
Extracting solution pH is adjusted to neutrality, and marine algae extract is made after being finally freeze-dried.
Embodiment 9
A kind of EDTA- citric acid antigens reparation liquid, the EDTA- citric acid antigens per 1000mL repair liquid and include：
Tris-Base 1.2114g, EDTA 0.37224g, monohydrate potassium 0.378252g, trisodium citrate dihydrate
The single of 2.411784g, 500 microlitres, glycerine 150mL, marine algae extract 1g of Tween-20 and surplus steams water.
Preparation method is as follows：Tris-Base, EDTA, monohydrate potassium, trisodium citrate dihydrate are weighed according to the ratio, are used
After single steaming water dissolution Tris-Base, EDTA, monohydrate potassium, trisodium citrate dihydrate, the glycerine of formula ratio is added, completely
Marine algae extract and Tween-20 are added after dissolving, are finally settled to rated capacity with single water that steams.
The preparation method of the marine algae extract is：By the seaweed after cleaning and water in mass ratio 3:10 mixing, are used in combination and beat
Seaweed slurry is made in pulp grinder；The hydrochloric acid solution by volume 1 for being 4 by seaweed slurry and pH value:1.5 mixing, obtain mixed slurry,
Microwave radiation exaraction is carried out to mixed slurry, wherein extraction conditions are：80 DEG C, time 7.5min of temperature, microwave power 900W are micro-
Wave frequency rate 2450MHZ.It is filtered to remove solid after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 105 DEG C, the ring of 6MPa
Curing sterilizing 1h under border；Seaweed extracted liquor after sterilizing to curing is filtered removal crude protein；Filtered seaweed is extracted
Liquid pH is adjusted to neutrality, and marine algae extract is made after being finally freeze-dried.Raw materials used in the present invention, equipment, if without especially saying
It is bright, it is the common raw material, equipment of this field；Method therefor in the present invention is unless otherwise noted the routine side of this field
Method.
The above is only presently preferred embodiments of the present invention, is not imposed any restrictions to the present invention, every according to the present invention
Technical spirit still falls within the technology of the present invention side to any simple modification, change and equivalent transformation made by above example
The protection domain of case.
Claims (6)
1. a kind of EDTA- citric acid antigens repair liquid, which is characterized in that the EDTA- citric acid antigens per 1000mL are repaired in liquid
Including：Tris-Base 1.20-1.25g, EDTA 0.35-0.40g, monohydrate potassium 0.35-0.40g, two citric acid monohydrates
Sodium 2.40-2.42g, 400-600 microlitres of Tween-20, triethanolamine or ethylene glycol or trihydroxylic alcohol 50-500mL, marine algae extract
0.5-1.5g and the single of surplus steam water.
2. a kind of EDTA- citric acid antigens as described in claim 1 repair liquid, which is characterized in that the EDTA- lemons per 1000mL
Lemon acid antigen retrieval buffers include：Tris-Base 1.2114g, EDTA 0.37224g, monohydrate potassium 0.378252g, two
Citric acid monohydrate sodium 2.411784g, 500 microlitres of Tween-20, triethanolamine or ethylene glycol or trihydroxylic alcohol 150mL, seaweed extraction
Object 1g and the single of surplus steam water.
3. a kind of EDTA- citric acid antigens as claimed in claim 2 repair liquid, which is characterized in that the EDTA- lemons per 1000mL
Lemon acid antigen retrieval buffers include：Tris-Base 1.2114g, EDTA 0.37224g, monohydrate potassium 0.378252g, two
The single of citric acid monohydrate sodium 2.411784g, 500 microlitres, triethanolamine 150mL, marine algae extract 1g of Tween-20 and surplus steams
Water.
4. a kind of EDTA- citric acid antigens as described in one of claim 1-3 repair liquid, it is characterised in that preparation method is such as
Under：Tris-Base, EDTA, monohydrate potassium and trisodium citrate dihydrate are weighed according to the ratio, steam water dissolution Tris- with single
After Base, EDTA, monohydrate potassium and trisodium citrate dihydrate, the triethanolamine or ethylene glycol or ternary of formula ratio is added
Alcohol adds marine algae extract and Tween-20 after being completely dissolved, be finally settled to rated capacity with single water that steams.
5. a kind of EDTA- citric acid antigens as described in one of claim 1-3 repair liquid, which is characterized in that the seaweed carries
The preparation method of object is taken to be：By the seaweed after cleaning and water 2-4 in mass ratio:10 mixing, are used in combination beater that seaweed slurry is made；
By hydrochloric acid solution that seaweed slurry and pH value are 3-5 by volume 1:1-2 is mixed, and obtains mixed slurry, is carried out to mixed slurry
Microwave radiation exaraction is filtered to remove solid after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 100-110 DEG C, 4-
Curing sterilizing 0.5-1.5h in the environment of 8MPa；Seaweed extracted liquor after sterilizing to curing is filtered removal crude protein；Incited somebody to action
Seaweed extracted liquor pH after filter is adjusted to neutrality, and marine algae extract is made after being finally freeze-dried.
6. a kind of EDTA- citric acid antigens as claimed in claim 5 repair liquid, which is characterized in that the microwave radiation exaraction
Condition be：70-90 DEG C of temperature, time 5-10min, microwave power 800-1000W, microwave frequency 2450MHZ.
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| CN109827821B (en) * | 2019-03-14 | 2021-11-09 | 武汉原谷生物科技有限责任公司 | Non-xylene dewaxing repairing agent for paraffin section and preparation method thereof |
| CN113495138A (en) * | 2021-06-22 | 2021-10-12 | 南京弗瑞思生物科技有限公司 | Tissue slice antigen repair liquid and use method thereof |
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