CN106053172A - EDTA antigen retrieval liquid - Google Patents
EDTA antigen retrieval liquid Download PDFInfo
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- CN106053172A CN106053172A CN201610385943.XA CN201610385943A CN106053172A CN 106053172 A CN106053172 A CN 106053172A CN 201610385943 A CN201610385943 A CN 201610385943A CN 106053172 A CN106053172 A CN 106053172A
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- Prior art keywords
- edta
- antigen retrieval
- retrieval buffers
- tris
- base
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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 92
- 239000000427 antigen Substances 0.000 title claims abstract description 80
- 102000036639 antigens Human genes 0.000 title claims abstract description 80
- 108091007433 antigens Proteins 0.000 title claims abstract description 80
- 239000007788 liquid Substances 0.000 title abstract description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 48
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims abstract description 45
- 239000007983 Tris buffer Substances 0.000 claims abstract description 38
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims abstract description 38
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 claims abstract description 28
- 238000010025 steaming Methods 0.000 claims abstract description 11
- 239000000872 buffer Substances 0.000 claims description 62
- 241000195474 Sargassum Species 0.000 claims description 45
- 241001474374 Blennius Species 0.000 claims description 29
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 29
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 29
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 29
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 21
- 238000002360 preparation method Methods 0.000 claims description 14
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 12
- 230000005070 ripening Effects 0.000 claims description 12
- 239000002002 slurry Substances 0.000 claims description 12
- 230000001954 sterilising effect Effects 0.000 claims description 12
- 238000001914 filtration Methods 0.000 claims description 11
- 239000011268 mixed slurry Substances 0.000 claims description 11
- 238000000605 extraction Methods 0.000 claims description 10
- 239000000243 solution Substances 0.000 claims description 10
- 238000004090 dissolution Methods 0.000 claims description 8
- 238000004140 cleaning Methods 0.000 claims description 7
- 230000005855 radiation Effects 0.000 claims description 7
- 235000019750 Crude protein Nutrition 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 6
- 230000007935 neutral effect Effects 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 4
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 11
- 238000003364 immunohistochemistry Methods 0.000 abstract description 8
- 238000004043 dyeing Methods 0.000 abstract description 6
- 230000002055 immunohistochemical effect Effects 0.000 abstract 1
- 230000008595 infiltration Effects 0.000 abstract 1
- 238000001764 infiltration Methods 0.000 abstract 1
- 210000001519 tissue Anatomy 0.000 description 18
- 230000008439 repair process Effects 0.000 description 14
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 12
- 238000000034 method Methods 0.000 description 9
- 238000010009 beating Methods 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
- 239000000203 mixture 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
- 238000002156 mixing Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000002904 solvent 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
- 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
- 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
- 230000000093 cytochemical effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- ZZVUWRFHKOJYTH-UHFFFAOYSA-N diphenhydramine Chemical compound C=1C=CC=CC=1C(OCCN(C)C)C1=CC=CC=C1 ZZVUWRFHKOJYTH-UHFFFAOYSA-N 0.000 description 1
- 239000000975 dye Substances 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
- 210000004877 mucosa 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 invention relates to the technical field of immunohistochemistry and discloses EDTA antigen retrieval liquid. Each 1,000 mL of EDTA antigen retrieval liquid is prepared from 1.20-1.25 g of Tris-Base, 0.35-0.40 g of EDTA, 400-600 microliters of twain-20, 50-500 mL of triethanolamine or ethanediol or triethanolamine and the balance single steaming water. The EDTA antigen retrieval liquid can achieve infiltration, is good in retrieval effect and can retrieve tissue slices with a little amount, the retrieved tissue slices can be dyed easily, and the dyeing effect is good. The retrieval liquid is not prone to volatilization and can be used in cooperation with a full-automatic immunohistochemical instrument.
Description
Technical field
The present invention relates to immunohistochemistry technology field, particularly relate to a kind of EDTA antigen retrieval buffers.
Background technology
Immunohistochemistry technology is the detection technique that pathological diagnosis in recent years develops rapidly, as a kind of important pathology
Aided diagnosis method, is increasingly subject to pay attention in clinical application.Along with the application of immunohistochemistry autostainer, right
Requirement in dyeing quality improves the most day by day.Wherein, antigen retrieval has of crucial importance during whole 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 himself
Structure feature, it is less for the container volume holding repair liquid, it is impossible to as repaired by hand, tissue slice is fully immersed in
Condition in repair liquid, and immunohistochemistry autostainer needs when antigen retrieval on higher temperature (100 DEG C of left sides
Right) under carry out, therefore repair liquid is readily volatilized, further reduces the volume of repair liquid.Thus, how in full-automatic immune group
Completing normal antigen retrieval on change dyeing instrument is the difficult problem that immunohistochemistry autostainer must solve.
But owing to current immunohistochemistry autostainer is domestic not yet common, few people use, therefore nobody
Notice above-mentioned technical problem.The most external method solving above-mentioned technical problem is typically to control the environment of antigen retrieval, as
Control atmospheric pressure, ambient temperature etc., these solution complicated conditions, be difficult to reality application.Also do not find by changing at present
Enter antigen retrieval buffers to solve the report of above-mentioned technical problem.
Current antigen retrieval buffers has a large amount in variety, such as EDTA antigen retrieval buffers, citric acid antigen repair liquid and EDTA-
Citric acid compounds antigen retrieval buffers etc..The Chinese patent of Application No. 201180024136.0 discloses a kind of antigen retrieval buffers
With antigen retrieval method, the antigen retrieval in the cytochemical staining procedure of immuning tissue and cell.Described antigen retrieval buffers is
Itaconic acid, itaconic anhydride or citraconic acid aqueous solution.Described antigen retrieval method is for using above-mentioned antigen retrieval buffers to tissue slice
Carry out high temperature or High Temperature High Pressure reparation.It is high that the antigen retrieval buffers provided and antigen retrieval method have dyeing quality, reproducibility,
Safety, the advantage of good stability.
The solvent of above-mentioned antigen retrieval buffers is water, if be applied on immunohistochemistry autostainer, on the one hand, this resists
Former repair liquid needs repairing effect in the case of consumption is big better, it is impossible to cannot be by tissue slice thorough impregnation at less consumption
In the case of complete preferably to repair；On the other hand, its boiling point is low, the most readily volatilized, is easily caused tissue and cuts
Sheet generation mummification situation, and once there is mummification, the antigen of tissue slice is changing to irreversible destruction and cannot repair.
Summary of the invention
In order to solve above-mentioned technical problem, the invention provides a kind of EDTA antigen retrieval buffers.The EDTA antigen of the present invention
Repair liquid infiltrates, repairing effect is good, it is possible to repairing tissue slice under less consumption, after reparation, tissue slice is easy
Dyeing, Color is good.And repair liquid is the most volatile, it is possible to coordinate full-automatic SABC instrument to use.
The concrete technical scheme of the present invention is: a kind of EDTA antigen retrieval buffers, in the EDTA antigen retrieval buffers of every 1000mL
Including: Tris-Base 1.20-1.25g, EDTA 0.35-0.40g, tween 20 400-600 microlitre, triethanolamine or second two
The list of alcohol or trihydroxylic alcohol 50-500mL and surplus steams water.
In the antigen retrieval buffers of the present invention, using EDTA as main repairing activity composition, and with triethanolamine or
Ethylene glycol or trihydroxylic alcohol steam water as mixed solvent with list.Wherein, triethanolamine or ethylene glycol or the addition of trihydroxylic alcohol, it is possible to carry
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 point
Agent is compared, and its advantage is that the impact on the active component in antigen retrieval buffers is less, and preferable with the affinity of tissue slice, poison
Property low, the reparation of tissue slice will not be caused too much impact, wherein trihydroxylic alcohol, the effect of triethylamine alcohol are better than ethylene glycol.Tell
Temperature-20 plays emulsification, it is possible to increase the compatibility of each material in antigen retrieval buffers.
As preferably, the EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base 1.2114g, EDTA
0.37224g, tween 20 500 microlitre, the list of triethanolamine or ethylene glycol or trihydroxylic alcohol 150mL and surplus steams water.
As preferably, the EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base 1.2114g, EDTA
0.37224g, tween 20 500 microlitre, the list of triethanolamine 150mL and surplus steams water.
As preferably, the EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base 1.20-1.25g, EDTA
0.35-0.40g, tween 20 400-600 microlitre, triethanolamine or ethylene glycol or trihydroxylic alcohol 50-500mL, Sargassum extract
The list of 0.5-1.5g and surplus steams water.
As preferably, the EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base 1.2114g, EDTA
0.37224g, tween 20 500 microlitre, triethanolamine or ethylene glycol or trihydroxylic alcohol 150mL, Sargassum extract 1g and the list of surplus
Steam water.
As preferably, the EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base 1.2114g, EDTA
0.37224g, tween 20 500 microlitre, triethanolamine 150mL, the list of Sargassum extract 1g and surplus steams water.
As preferably, the preparation method of described EDTA antigen retrieval buffers is as follows:
Weigh Tris-Base and EDTA by proportioning, after single steaming water dissolution Tris-Base and EDTA, add three second of formula ratio
Hydramine or ethylene glycol or trihydroxylic alcohol, add tween 20, be finally settled to rated capacity with single steaming water after being completely dissolved.
As preferably, the preparation method of described EDTA antigen retrieval buffers also can be as follows: by proportioning weigh Tris-Base and
EDTA, after single steaming water dissolution Tris-Base and EDTA, the triethanolamine of addition formula ratio or ethylene glycol or trihydroxylic alcohol, completely
Add Sargassum extract and tween 20 after dissolving, be finally settled to rated capacity with single water that steams.
As preferably, the preparation method of described Sargassum extract is: the Sargassum after cleaning and water 2-4:10 in mass ratio
Mixing, and make seaweed slurry with beater；Seaweed slurry is mixed with the hydrochloric acid solution that pH value is 3-5 1:1-2 by volume,
Obtain mixed slurry, mixed slurry is carried out microwave radiation exaraction, be filtered to remove solids after extraction and obtain seaweed extracted liquor, right
Seaweed extracted liquor at 100-110 DEG C, ripening sterilizing 0.5-1.5h in the environment of 4-8MPa, part can be made in maturing process many
Sugar degraded；Seaweed extracted liquor after ripening sterilizing is filtered the crude protein removing degeneration；Sargassum after filtering is extracted
Liquid pH regulator, to neutral, prepares Sargassum extract after last lyophilization.
In order to further optimize, the antigen retrieval buffers of the present invention also adds Sargassum extract.The present invention prepares
Sargassum extract contains the materials such as natural alginate and natural polysaccharide, and biological mildness is good, with tissue slice affinity
Good.A small amount of Sargassum extract viscosity that liquid can be made certain soluble in water, lock water is effective, it is possible to prevent antigen further
Solvent volatilization in repair liquid.Additionally, Sargassum extract to the repairing activity composition in antigen retrieval buffers can physical bond, Sargassum
After extract contacts with tissue slice, it is possible to form one layer of thin mucosa on tissue slice surface, so that in connection
Repairing activity composition can fully contact with tissue slice, in the case of less consumption antigen retrieval buffers, improves repairing effect.
But the used in amounts of Sargassum extract if consumption is too much, can cause antigen retrieval fluid viscosity too high, mobility strictly control
The poorest, and be combined excessively with tissue slice, it is difficult to follow-up cleaning.
As preferably, the condition of described microwave radiation exaraction is: temperature 70-90 DEG C, time 5-10min, microwave power
800-1000W, microwave frequency 2450MHZ.
As preferably, described trihydroxylic alcohol can be glycerol.
It is compared with the prior art, the invention has the beneficial effects as follows:
The EDTA antigen retrieval buffers of the present invention infiltrates, repairing effect is good, it is possible to repair tissue slice under less consumption
Multiple.Repair liquid needed for averagely every tissue slice only needs 100-150uL.
After reparation, tissue slice easily dyes, and Color is good.
And repair liquid is the most volatile, it is possible to coordinate full-automatic SABC instrument to use.
Detailed description of the invention
Below in conjunction with embodiment, the invention will be further described.
Embodiment 1
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base 1.2114g, EDTA
0.37224g, tween 20 500 microlitre, the list of triethanolamine 150mL and surplus steams water.
Preparation method is as follows: weigh Tris-Base and EDTA by proportioning, after single steaming water dissolution Tris-Base and EDTA,
Add the triethanolamine of formula ratio, add tween 20 after being completely dissolved, be finally settled to rated capacity with single water that steams.
Embodiment 2
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base 1.2114g, EDTA
0.37224g, tween 20 500 microlitre, the list of ethylene glycol 325mL and surplus steams water.
Preparation method is as follows: weigh Tris-Base and EDTA by proportioning, after single steaming water dissolution Tris-Base and EDTA,
Add the ethylene glycol of formula ratio, add tween 20 after being completely dissolved, be finally settled to rated capacity with single water that steams.
Embodiment 3
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base 1.20g, EDTA
0.35g, tween 20 400 microlitre, the list of triethanolamine 50mL and surplus steams water.
Embodiment 4
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base 1.25g, EDTA
0.40g, tween 20 600 microlitre, the list of triethanolamine 500mL and surplus steams water.
Embodiment 5
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base 1.2114g, EDTA
0.37224g, tween 20 500 microlitre, the list of glycerol 300mL and surplus steams water.
Embodiment 6
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base 1.2114g,
EDTA 0.37224g, tween 20 500 microlitre, triethanolamine 150mL, the list of Sargassum extract 1g and surplus steams water.
Preparation method is as follows: weigh Tris-Base and EDTA by proportioning, after single steaming water dissolution Tris-Base and EDTA,
Add the triethanolamine of formula ratio, add Sargassum extract and tween 20 after being completely dissolved, be finally settled to volume with single water that steams
Constant volume.
The preparation method of described Sargassum extract is: the Sargassum after cleaning mixes with water 3:10 in mass ratio, and with beating
Pulp grinder makes seaweed slurry；Seaweed slurry is mixed with the hydrochloric acid solution that pH value is 4 1:1.5 by volume, obtains mixed slurry,
Mixed slurry carries out microwave radiation exaraction, and wherein extraction conditions is: temperature 80 DEG C, and time 7.5min, microwave power 900W are micro-
Wave frequency 2450MHZ.It is filtered to remove solids after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 105 DEG C, the ring of 6MPa
Ripening sterilizing 1h under border；Carry out the seaweed extracted liquor after ripening sterilizing filtering and remove crude protein；Sargassum after filtering is extracted
Liquid pH regulator, to neutral, prepares Sargassum extract after last lyophilization.
Embodiment 7
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers of every 1000mL includes: the EDTA antigen of every 1000mL is repaiied
Multiple liquid includes: Tris-Base 1.2114g, EDTA 0.37224g, tween 20 500 microlitre, triethanolamine 150mL, Sargassum
The list of extract 0.5g and surplus steams water.
The preparation method of described Sargassum extract is: the Sargassum after cleaning mixes with water 2:10 in mass ratio, and with beating
Pulp grinder makes seaweed slurry；Seaweed slurry is mixed with the hydrochloric acid solution that pH value is 3-5 1:1 by volume, obtains mixed slurry,
Mixed slurry carries out microwave radiation exaraction, and wherein extraction conditions is: temperature 70 C, time 10min, and microwave power 800W is micro-
Wave frequency 2450MHZ.It is filtered to remove solids after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 100 DEG C, 8MPa's
Ripening sterilizing 0.5h under environment；Carry out the seaweed extracted liquor after ripening sterilizing filtering and remove crude protein；Sargassum after filtering
Extracting solution pH regulator, to neutral, prepares Sargassum extract after last lyophilization.
Embodiment 8
The EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base 1.2114g, EDTA 0.37224g, tween 20
500 microlitres, triethanolamine 150mL, the list of Sargassum extract 1.5g and surplus steams water.
The preparation method of described Sargassum extract is: the Sargassum after cleaning mixes with water 4:10 in mass ratio, and with beating
Pulp grinder makes seaweed slurry；Seaweed slurry is mixed with the hydrochloric acid solution that pH value is 3-5 1:2 by volume, obtains mixing slurry
Material, carries out microwave radiation exaraction to mixed slurry, and wherein extraction conditions is: temperature 90 DEG C, time 5min, microwave power 1000W,
Microwave frequency 2450MHZ.It is filtered to remove solids after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 110 DEG C, 4MPa's
Ripening sterilizing 1.5h under environment；Carry out the seaweed extracted liquor after ripening sterilizing filtering and remove crude protein；Sargassum after filtering
Extracting solution pH regulator, to neutral, prepares Sargassum extract after last lyophilization.
Embodiment 9
A kind of EDTA antigen retrieval buffers, the EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base 1.2114g, EDTA
0.37224g, tween 20 500 microlitre, glycerol 150mL, the list of Sargassum extract 1g and surplus steams water.
Preparation method is as follows: weigh Tris-Base and EDTA by proportioning, after single steaming water dissolution Tris-Base and EDTA,
Add the glycerol of formula ratio, add Sargassum extract and tween 20 after being completely dissolved, be finally settled to specified appearance with single water that steams
Amount.
The preparation method of described Sargassum extract is: the Sargassum after cleaning mixes with water 3:10 in mass ratio, and with beating
Pulp grinder makes seaweed slurry；Seaweed slurry is mixed with the hydrochloric acid solution that pH value is 4 1:1.5 by volume, obtains mixed slurry,
Mixed slurry carries out microwave radiation exaraction, and wherein extraction conditions is: temperature 80 DEG C, and time 7.5min, microwave power 900W are micro-
Wave frequency 2450MHZ.It is filtered to remove solids after extraction and obtains seaweed extracted liquor, to seaweed extracted liquor at 105 DEG C, the ring of 6MPa
Ripening sterilizing 1h under border；Carry out the seaweed extracted liquor after ripening sterilizing filtering and remove crude protein；Sargassum after filtering is extracted
Liquid pH regulator, to neutral, prepares Sargassum extract after last lyophilization.Raw materials used, equipment in the present invention, if without saying especially
Bright, it is the conventional raw material of this area, equipment；Method therefor in the present invention, unless otherwise noted, is the routine side of this area
Method.
The above, be only presently preferred embodiments of the present invention, not impose any restrictions the present invention, every according to the present invention
Any simple modification, change and the equivalent transformation that above example is made by technical spirit, all still falls within the technology of the present invention side
The protection domain of case.
Claims (10)
1. an EDTA antigen retrieval buffers, it is characterised in that: the EDTA antigen retrieval buffers of every 1000mL includes: Tris-Base
1.20-1.25g, EDTA 0.35-0.40g, tween 20 400-600 microlitre, triethanolamine or ethylene glycol or trihydroxylic alcohol 50-
The list of 500mL and surplus steams water.
2. a kind of EDTA antigen retrieval buffers as claimed in claim 1, it is characterised in that the EDTA antigen retrieval buffers of every 1000mL
Include: Tris-Base 1.2114g, EDTA 0.37224g, tween 20 500 microlitre, triethanolamine or ethylene glycol or ternary
The list of alcohol 150mL and surplus steams water.
3. a kind of EDTA antigen retrieval buffers as claimed in claim 2, it is characterised in that the EDTA antigen retrieval buffers of every 1000mL
Include: Tris-Base 1.2114g, EDTA 0.37224g, tween 20 500 microlitre, triethanolamine 150mL and surplus
Single steaming water.
4. a kind of EDTA antigen retrieval buffers as claimed in claim 1, it is characterised in that the EDTA antigen retrieval buffers of every 1000mL
Include: Tris-Base 1.20-1.25g, EDTA 0.35-0.40g, tween 20 400-600 microlitre, triethanolamine or second
Glycol or trihydroxylic alcohol 50-500mL, the list of Sargassum extract 0.5-1.5g and surplus steams water.
5. a kind of EDTA antigen retrieval buffers as claimed in claim 4, it is characterised in that the EDTA antigen retrieval buffers of every 1000mL
Include: Tris-Base 1.2114g, EDTA 0.37224g, tween 20 500 microlitre, triethanolamine or ethylene glycol or ternary
Alcohol 150mL, the list of Sargassum extract 1g and surplus steams water.
6. a kind of EDTA antigen retrieval buffers as claimed in claim 5, it is characterised in that the EDTA antigen retrieval buffers of every 1000mL
Including: Tris-Base 1.2114g, EDTA 0.37224g, tween 20 500 microlitre, triethanolamine 150mL, Sargassum is extracted
The list of thing 1g and surplus steams water.
7. 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: by proportioning
Weigh Tris-Base and EDTA, after single steaming water dissolution Tris-Base and EDTA, add triethanolamine or the second two of formula ratio
Alcohol or trihydroxylic alcohol, add tween 20, be finally settled to rated capacity with single water that steams after being completely dissolved.
8. a kind of EDTA antigen retrieval buffers as described in one of claim 4-6, it is characterised in that preparation method is as follows: by proportioning
Weigh Tris-Base and EDTA, after single steaming water dissolution Tris-Base and EDTA, add triethanolamine or the second two of formula ratio
Alcohol or trihydroxylic alcohol, add Sargassum extract and tween 20, be finally settled to rated capacity with single water that steams after being completely dissolved.
9. a kind of EDTA antigen retrieval buffers as described in one of claim 4-6, it is characterised in that the system of described Sargassum extract
Preparation Method is: the Sargassum after cleaning mixes with water 2-4:10 in mass ratio, and makes seaweed slurry with beater；Sargassum is starched
Material mixes with the hydrochloric acid solution that pH value is 3-5 1:1-2 by volume, obtains mixed slurry, mixed slurry is carried out microwave-assisted
Extract, be filtered to remove solids after extraction and obtain seaweed extracted liquor, to seaweed extracted liquor at 100-110 DEG C, the environment of 4-8MPa
Lower ripening sterilizing 0.5-1.5h；Carry out the seaweed extracted liquor after ripening sterilizing filtering and remove crude protein；Sargassum after filtering
Extracting solution pH regulator, to neutral, prepares Sargassum extract after last lyophilization.
10. a kind of EDTA antigen retrieval buffers as claimed in claim 9, it is characterised in that the condition of described microwave radiation exaraction
For: temperature 70-90 DEG C, time 5-10min, microwave power 800-1000W, microwave frequency 2450MHZ.
Priority Applications (1)
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