CN106053172B - A kind of EDTA antigen retrieval buffers - Google Patents
A kind of EDTA antigen retrieval buffers Download PDFInfo
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- CN106053172B CN106053172B CN201610385943.XA CN201610385943A CN106053172B CN 106053172 B CN106053172 B CN 106053172B CN 201610385943 A CN201610385943 A CN 201610385943A CN 106053172 B CN106053172 B CN 106053172B
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- retrieval buffers
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- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 title claims abstract description 80
- 239000000427 antigen Substances 0.000 title claims abstract description 72
- 102000036639 antigens Human genes 0.000 title claims abstract description 72
- 108091007433 antigens Proteins 0.000 title claims abstract description 72
- 239000000872 buffer Substances 0.000 title claims abstract description 59
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 43
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims abstract description 42
- 239000007983 Tris buffer Substances 0.000 claims abstract description 33
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims abstract description 33
- 229920001213 Polysorbate 20 Polymers 0.000 claims abstract description 25
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims abstract description 25
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims abstract description 25
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 claims abstract description 24
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 18
- 241001474374 Blennius Species 0.000 claims description 44
- 241000195493 Cryptophyta Species 0.000 claims description 30
- 238000002360 preparation method Methods 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
- 239000002002 slurry Substances 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 11
- 230000005855 radiation Effects 0.000 claims description 8
- 238000004140 cleaning Methods 0.000 claims description 7
- 238000004090 dissolution Methods 0.000 claims description 7
- 235000019750 Crude protein Nutrition 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 238000010025 steaming Methods 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 2
- 239000007788 liquid Substances 0.000 abstract description 17
- 230000000694 effects Effects 0.000 abstract description 11
- 238000003364 immunohistochemistry Methods 0.000 abstract description 11
- 230000008439 repair process Effects 0.000 abstract description 8
- 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
- 238000000034 method Methods 0.000 description 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 8
- 235000011187 glycerol Nutrition 0.000 description 4
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 239000004615 ingredient Substances 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
- 239000002994 raw material Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 210000001519 tissue Anatomy 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
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 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
- 239000003795 chemical substances by application Substances 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
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 238000005470 impregnation Methods 0.000 description 1
- 230000002427 irreversible effect Effects 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
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 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
- 230000009466 transformation Effects 0.000 description 1
Classifications
-
- 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
-
- 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
Abstract
The present invention relates to immunohistochemistry technology fields, disclose a kind of EDTA antigen retrieval buffers, and the EDTA antigen retrieval buffers per 1000mL include：Tris-Base 1.20-1.25g, EDTA 0.35-0.40g, 400-600 microlitres of Tween-20, triethanolamine or ethylene glycol or trihydroxylic alcohol 50-500mL and the single of surplus steam water.The EDTA antigen retrieval buffers infiltration of the present invention, repairing effect are good, can be repaired to histotomy under less dosage, and 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 field more particularly to a kind of EDTA antigen retrieval buffers.
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 antigen retrieval buffers.The EDTA antigens of the present invention
The infiltration of reparation liquid, repairing effect are good, can be repaired to histotomy under less dosage, histotomy is easy after reparation
Dyeing, coloring are good.And it is not volatile to repair liquid, and full-automatic immunohistochemistry instrument can be coordinated to use.
The specific technical solution of the present invention is：A kind of EDTA antigen retrieval buffers, often in the EDTA antigen retrieval buffers of 1000mL
Including：Tris-Base 1.20-1.25g, EDTA 0.35-0.40g, 400-600 microlitres of Tween-20, triethanolamine or second two
Alcohol or trihydroxylic alcohol 50-500mL and the single of surplus steam water.
In the antigen retrieval buffers of the present invention, using EDTA as main repairing activity ingredient, and with triethanolamine or
Ethylene glycol or trihydroxylic alcohol steam water as mixed solvent with list.Wherein, the addition of triethanolamine or ethylene glycol or trihydroxylic alcohol, Neng Gouti
The boiling point of high antigen retrieval buffers, prevents it from excessively volatilizing.And triethanolamine or ethylene glycol or trihydroxylic alcohol are molten with other high points
Agent is compared, and advantage is that the influence to the active constituent in antigen retrieval buffers is smaller, and preferable with the compatibility of histotomy, poison
Property it is low, the reparation of histotomy will not be caused excessively to influence, wherein trihydroxylic alcohol, triethylamine alcohol effect be better than ethylene glycol.It spits
Temperature -20 plays emulsification, can improve the compatibility of each substance in antigen retrieval buffers.
Preferably, the EDTA antigen retrieval buffers per 1000mL include：Tris-Base 1.2114g, EDTA
0.37224g, 500 microlitres of Tween-20, triethanolamine or ethylene glycol or trihydroxylic alcohol 150mL and the single of surplus steam water.
Preferably, the EDTA antigen retrieval buffers per 1000mL include：Tris-Base 1.2114g, EDTA
The single of 0.37224g, 500 microlitres of Tween-20, triethanolamine 150mL and surplus steams water.
Preferably, the EDTA antigen retrieval buffers per 1000mL include：Tris-Base 1.20-1.25g, EDTA
0.35-0.40g, 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.
Preferably, the EDTA antigen retrieval buffers per 1000mL include：Tris-Base 1.2114g, EDTA
0.37224g, 500 microlitres of Tween-20, triethanolamine or ethylene glycol or trihydroxylic alcohol 150mL, the list of marine algae extract 1g and surplus
Steam water.
Preferably, the EDTA antigen retrieval buffers per 1000mL include：Tris-Base 1.2114g, EDTA
The single of 0.37224g, 500 microlitres, triethanolamine 150mL, marine algae extract 1g of Tween-20 and surplus steams water.
Preferably, the preparation method of the EDTA antigen retrieval buffers is as follows：
Tris-Base and EDTA are weighed according to the ratio, and after single steaming water dissolution Tris-Base and EDTA, formula ratio is added
Triethanolamine or ethylene glycol or trihydroxylic alcohol, add Tween-20 after being completely dissolved, be finally settled to rated capacity with single water that steams.
Preferably, the preparation method of the EDTA antigen retrieval buffers also can be as follows：Weigh according to the ratio Tris-Base and
EDTA, with it is single steam water dissolution Tris-Base and EDTA after, the triethanolamine or ethylene glycol or trihydroxylic alcohol 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.
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 present invention EDTA antigen retrieval buffers infiltration, repairing effect it is good, can under less dosage to histotomy into
Row 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.
Embodiment 1
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers per 1000mL include：Tris-Base 1.2114g,
The single of EDTA 0.37224g, 500 microlitres of Tween-20, triethanolamine 150mL and surplus steams water.
Preparation method is as follows：Tris-Base and EDTA are weighed according to the ratio, after single steaming water dissolution Tris-Base and EDTA,
The triethanolamine of formula ratio is added, Tween-20 is added after being completely dissolved, is finally settled to rated capacity with single water that steams.
Embodiment 2
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers per 1000mL include：Tris-Base 1.2114g,
The single of EDTA 0.37224g, 500 microlitres of Tween-20, ethylene glycol 325mL and surplus steams water.
Preparation method is as follows：Tris-Base and EDTA are weighed according to the ratio, after single steaming water dissolution Tris-Base and EDTA,
The ethylene glycol of formula ratio is added, Tween-20 is added after being completely dissolved, is finally settled to rated capacity with single water that steams.
Embodiment 3
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers per 1000mL include：Tris-Base 1.20g,
The single of EDTA 0.35g, 400 microlitres of Tween-20, triethanolamine 50mL and surplus steams water.
Embodiment 4
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers per 1000mL include：Tris-Base 1.25g,
The single of EDTA 0.40g, 600 microlitres of Tween-20, triethanolamine 500mL and surplus steams water.
Embodiment 5
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers per 1000mL include：Tris-Base 1.2114g,
The single of EDTA 0.37224g, 500 microlitres of Tween-20, glycerine 300mL and surplus steams water.
Embodiment 6
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers per 1000mL include： Tris-Base
The single of 1.2114g, EDTA 0.37224g, 500 microlitres, triethanolamine 150mL, marine algae extract 1g of Tween-20 and surplus steams
Water.
Preparation method is as follows：Tris-Base and EDTA are weighed according to the ratio, after single steaming water dissolution Tris-Base and EDTA,
The triethanolamine of formula ratio is added, marine algae extract and Tween-20 are added after being completely dissolved, is finally settled to volume with single water that steams
Constant volume.
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 antigen retrieval buffers, the EDTA antigen retrieval buffers per 1000mL include：EDTA per 1000mL is anti-
Original repairs liquid：Tris-Base 1.2114g, EDTA 0.37224g, 500 microlitres, triethanolamine 150mL of Tween-20,
Marine algae extract 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
Often the EDTA antigen retrieval buffers of 1000mL include：Tris-Base 1.2114g, EDTA 0.37224g, tween-
The single of 20 500 microlitres, triethanolamine 150mL, marine algae extract 1.5g and surplus steams 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 antigen retrieval buffers, the EDTA antigen retrieval buffers per 1000mL include：Tris-Base 1.2114g,
The single of EDTA 0.37224g, 500 microlitres, glycerine 150mL, marine algae extract 1g of Tween-20 and surplus steams water.
Preparation method is as follows：Tris-Base and EDTA are weighed according to the ratio, after single steaming water dissolution Tris-Base and EDTA,
The glycerine of formula ratio is added, marine algae extract and Tween-20 are added after being completely dissolved, is finally settled to specified appearance with single water that steams
Amount.
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 antigen retrieval buffers, which is characterized in that often the EDTA antigen retrieval buffers of 1000mL include：Tris-Base
1.20-1.25g, EDTA 0.35-0.40g, 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 antigen retrieval buffers as described in claim 1, which is characterized in that the EDTA antigen retrieval buffers per 1000mL
Include：Tris-Base 1.2114g, EDTA 0.37224g, 500 microlitres of Tween-20, triethanolamine or ethylene glycol or ternary
Alcohol 150mL, marine algae extract 1g and the single of surplus steam water.
3. a kind of EDTA antigen retrieval buffers as claimed in claim 2, which is characterized in that the EDTA antigen retrieval buffers per 1000mL
Include：Tris-Base 1.2114g, EDTA 0.37224g, 500 microlitres, triethanolamine 150mL of Tween-20, seaweed extraction
Object 1g and the single of surplus steam water.
4. a kind of EDTA antigen retrieval buffers as described in one of claim 1-3, it is characterised in that preparation method is as follows：According to the ratio
Tris-Base and EDTA are weighed, after single steaming water dissolution Tris-Base and EDTA, the triethanolamine or second two of formula ratio is added
Alcohol or trihydroxylic alcohol add 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 antigen retrieval buffers as described in one of claim 1-3, which is characterized in that the system of the marine algae extract
Preparation Method is：By the seaweed after cleaning and water 2-4 in mass ratio:10 mixing, are used in combination beater that seaweed slurry is made；Seaweed is starched
Material and the hydrochloric acid solution by volume 1 that pH value is 3-5:1-2 is mixed, and obtains mixed slurry, and microwave radiation technology is carried out to mixed slurry
Extraction, is filtered to remove solid after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 100-110 DEG C, the environment of 4-8MPa
Lower curing sterilizing 0.5-1.5h；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.
6. a kind of EDTA antigen retrieval buffers as claimed in claim 5, which is characterized in that the condition of the microwave radiation exaraction
For：70-90 DEG C of temperature, time 5-10min, microwave power 800-1000W, microwave frequency 2450MHZ.
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| CN109975095A (en) * | 2019-02-22 | 2019-07-05 | 嘉兴雅康博医学检验所有限公司 | A kind of pretreatment liquid and pre-treating method of fluorescence in situ hybridization |
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