CN104808005B - A kind of method differentiating male epinephelus lanceolatus fish - Google Patents
A kind of method differentiating male epinephelus lanceolatus fish Download PDFInfo
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- CN104808005B CN104808005B CN201510179104.8A CN201510179104A CN104808005B CN 104808005 B CN104808005 B CN 104808005B CN 201510179104 A CN201510179104 A CN 201510179104A CN 104808005 B CN104808005 B CN 104808005B
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- 241001217936 Epinephelus lanceolatus Species 0.000 title claims abstract description 55
- 238000000034 method Methods 0.000 title claims abstract description 30
- 210000002966 serum Anatomy 0.000 claims abstract description 45
- MUMGGOZAMZWBJJ-DYKIIFRCSA-N Testostosterone Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 MUMGGOZAMZWBJJ-DYKIIFRCSA-N 0.000 claims abstract description 40
- 108010090932 Vitellogenins Proteins 0.000 claims abstract description 20
- 229960003604 testosterone Drugs 0.000 claims abstract description 20
- 210000004369 blood Anatomy 0.000 claims abstract description 9
- 239000008280 blood Substances 0.000 claims abstract description 9
- 238000005119 centrifugation Methods 0.000 claims abstract description 9
- 238000002965 ELISA Methods 0.000 claims abstract description 8
- 125000000468 ketone group Chemical group 0.000 claims abstract 4
- 239000000126 substance Substances 0.000 claims description 8
- 238000005259 measurement Methods 0.000 claims description 7
- 230000013011 mating Effects 0.000 claims description 5
- 238000003149 assay kit Methods 0.000 claims description 4
- 230000037396 body weight Effects 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 claims 2
- 108090000790 Enzymes Proteins 0.000 claims 2
- 210000002969 egg yolk Anatomy 0.000 claims 1
- 241000251468 Actinopterygii Species 0.000 abstract description 22
- 208000027418 Wounds and injury Diseases 0.000 abstract description 4
- 230000006378 damage Effects 0.000 abstract description 4
- 208000014674 injury Diseases 0.000 abstract description 4
- 239000013024 dilution buffer Substances 0.000 description 6
- 239000000872 buffer Substances 0.000 description 5
- 239000012895 dilution Substances 0.000 description 5
- 238000010790 dilution Methods 0.000 description 5
- 210000002149 gonad Anatomy 0.000 description 5
- 239000011534 wash buffer Substances 0.000 description 5
- 230000008901 benefit Effects 0.000 description 4
- 239000000700 radioactive tracer Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 description 3
- 241000321428 Epinephelus guttatus Species 0.000 description 3
- 238000009395 breeding Methods 0.000 description 3
- 229940088597 hormone Drugs 0.000 description 3
- 239000005556 hormone Substances 0.000 description 3
- 208000027877 Disorders of Sex Development Diseases 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 230000002710 gonadal effect Effects 0.000 description 2
- 201000005611 hermaphroditism Diseases 0.000 description 2
- 230000010196 hermaphroditism Effects 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 208000013327 true hermaphroditism Diseases 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 102000004277 11-beta-hydroxysteroid dehydrogenases Human genes 0.000 description 1
- 108090000874 11-beta-hydroxysteroid dehydrogenases Proteins 0.000 description 1
- WTPMRQZHJLJSBO-XQALERBDSA-N 11-oxotestosterone Chemical compound O=C1CC[C@]2(C)[C@H]3C(=O)C[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 WTPMRQZHJLJSBO-XQALERBDSA-N 0.000 description 1
- 241000489963 Anabantoidei Species 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 241000448486 Epinephelinae Species 0.000 description 1
- 241000357439 Epinephelus Species 0.000 description 1
- 241000917703 Leia Species 0.000 description 1
- 241000276618 Perciformes Species 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
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- 230000003292 diminished effect Effects 0.000 description 1
- 238000012850 discrimination method Methods 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 229940011871 estrogen Drugs 0.000 description 1
- 239000000262 estrogen Substances 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000009313 farming Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000007373 indentation Methods 0.000 description 1
- 230000000366 juvenile effect Effects 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 108010052522 livetin Proteins 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000010413 mother solution Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000031787 nutrient reservoir activity Effects 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000010172 protogyny Effects 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 210000001550 testis Anatomy 0.000 description 1
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- 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
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
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- 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
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
- G01N33/743—Steroid hormones
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Abstract
The invention discloses a kind of method differentiating male epinephelus lanceolatus fish, comprise the following steps：(1) select epinephelus lanceolatus fish：The epinephelus lanceolatus fish in selected reproduction season；(2) obtain epinephelus lanceolatus fish serum：Choose epinephelus lanceolatus fish tail venous blood, after centrifugation, obtain serum；(3) measure vitellogenin content in serum：Using the vitellogenin content in enzyme-linked immunosorbent assay serum：(4) 11 ketone group testosterone concentrations in serum are measured：Using 11 ketone group testosterone concentrations in enzyme-linked immunosorbent assay serum；(5) differentiate epinephelus lanceolatus fish milter：In conjunction with vitellogenin content in epinephelus lanceolatus fish serum and 11 ketone group testosterone concentrations, identify epinephelus lanceolatus fish milter.Rapidly and efficiently, the injury to fish is little for the method, and accuracy is high.
Description
Technical field
The invention belongs to cabrilla sex abnormality technical field and in particular to a kind of discriminating male epinephelus lanceolatus fish side
Method.
Background technology
Epinephelus lanceolatus fish is dragon wholesale, Perciformess, Anabantoidei, section, Epinephelinae again, and Epinepheluses are bodies in cabrilla
The maximum one kind of type.Epinephelus lanceolatus fish individuality is big, nutritious, delicious meat, fast growth, is the marine fish that people like
Treasure.In recent years because offshore is excessively captured, yield falls sharply, and supply falls short of demand in market.In coastal areas of southern China Guangdong, Hainan, Fujian, wide
The ground such as west epinephelus lanceolatus fish has become the main object of sea-farming, and economic worth is huge.
Epinephelus lanceolatus piscinity is ripe slow and existence reversal, belongs to protogyny, hermaphroditism Fish.Exactly
Because this characteristic of hermaphroditism, it is difficult to only pick out male parent population from external appearance characteristic, due to this therefore in selection-breeding parent fish
Reason leads to epinephelus lanceolatus fish parent fish to cultivate difficulty increasing, becomes a difficult problem of epinephelus lanceolatus fish breeding work.
At present, male 3 points of epinephelus lanceolatus fish Main Basiss are judged：Build and the size at age, see reproduction hole site, directly
Connect the method extracting gonad.Three kinds of methods respectively have pluses and minuses, and it is convenient that the size according to build and age judges, the time is short, operation
Simply, but this method accuracy is low, the impact of the subjectivity of people is too big, feeding method simultaneously, and growing environment affects on it to compare
Larger.See gonopore fast position, simple to operate, to fish fanout free region, but, non-reproduction period, gonopore often diminished slight indentation
Abdominal part is at this time to be difficult to judge male and female, and accuracy is low.By directly extracting the method judgment accuracy height of gonad, but
It is that the injury ratio to fish is larger, impact gonadal maturation even causes death, complex operation simultaneously, and workload is big, during judgement
Between long.
Vitellogenin (vitellogenin, Vtg) is specifically to be present in oviparity non-mammalian sexual maturity animal blood
A kind of albumen, be the precursor of livetin (Yolk protein).Participate in oviparous animal as a kind of important reproduction albumen
The important physiological process such as reproduction, growth.Vitellogenin is considered as a kind of preferable estrogen and oestrogen-like hormone mark.Logical
Often Vtg can only detect in the raun blood plasma of idiophase, and milter and juvenile fish in-vivo content are very small.11- ketone group testosterone
(11-ketotestosterone, 11-KT), is the important male steroid of one of Fish.It is transformed by testosterone, testosterone
It is changed into 11p- hydroxytestosterone, the latter forms 11-KT by 11beta-Hydroxysteroid dehydrogenase catalysis again, directly induce fish sperm
Generate.
In mating period, the content of male and female Fish both hormones has larger difference.Therefore, the present invention intends by detection
With compare content in serum for both hormones and then identify male epinephelus lanceolatus fish.
Content of the invention
It is an object of the invention to provide a kind of method differentiating male epinephelus lanceolatus fish, the method rapidly and efficiently, to fish
Injury little, accuracy is high.
The above-mentioned purpose of the present invention can be achieved through the following technical solutions：A kind of side differentiating male epinephelus lanceolatus fish
Method, comprises the following steps：
(1) select epinephelus lanceolatus fish：The epinephelus lanceolatus fish in selected reproduction season；
(2) obtain epinephelus lanceolatus fish serum：Choose epinephelus lanceolatus fish tail venous blood, after centrifugation, obtain serum；
(3) measure vitellogenin content in serum：Using the vitellogenin in enzyme-linked immunosorbent assay serum
Content：
(4) measure 11- ketone group testosterone concentration in serum：Using the 11- ketone group testis in enzyme-linked immunosorbent assay serum
Ketone content；
(5) differentiate epinephelus lanceolatus fish milter：In conjunction with vitellogenin content in epinephelus lanceolatus fish serum and 11- ketone group testosterone
Content, identifies epinephelus lanceolatus fish milter.
In the method for above-mentioned discriminating male epinephelus lanceolatus fish：
In step (1), selective body focuses on the epinephelus lanceolatus fish of the mating period of more than 20kg, and mating period is coastal in south
It is preferably 5 annual～August.
Preferably choose epinephelus lanceolatus fish tail venous blood 2～5mL in step (2), after centrifugation, obtain serum.
In step (2), during centrifugation, centrifuge speed is preferably 3500～5000rpm, and centrifuging temperature is preferably 4 DEG C, during centrifugation
Between be preferably 10～15min.
When adopting the vitellogenin content in enzyme-linked immunosorbent assay serum in step (3), it is preferred to use
The serum vitellogenin assay kit measurement of cayman chemical company, specific flow process is with reference to reagent
The operation instruction of box.
When adopting the 11- ketone group testosterone concentration in enzyme-linked immunosorbent assay serum in step (4), it is preferred to use
The serum 11- ketone group testosterone concentration of cayman chemical company measures kit measurement, and specific flow process is with reference to reagent
The operation instruction of box.
When vitellogenin content ＜ 0.9mg/mL and 11- ketone group testosterone concentration in epinephelus lanceolatus fish serum in step (5)
During ＞ 1ng/mL, differentiate as epinephelus lanceolatus fish milter.
The invention has the advantages that：The inventive method is contained by measuring vitellogenin in epinephelus lanceolatus fish serum simultaneously
Amount and 11- ketone group testosterone concentration carry out the discriminating of male epinephelus lanceolatus fish, and accuracy is high, simple, and fish body are injured little.
Specific embodiment
Embodiment 1
A kind of method of discriminating epinephelus lanceolatus fish milter that the present embodiment provides, comprises the following steps that：
(1) in the saddle of No. 21 more than the 20kg choosing 30 Dayawan Aquatic Products Experimental Center, Guangdong Prov.'s cultivation of in June, 2014
Band cabrilla, now from the male and female that can not identify these fishes in appearance, stamps electronic marker on every fish dorsal muscle,
The corresponding labelling number of the good every fish of record；
(2) adopt tail vein blood taking method, put in the centrifuge tube of 15mL with the blood that 50mL syringe extracts 2mL, will fill
The centrifuge tube having blood is placed on 4 DEG C of refrigerator overnight, and 4 DEG C on centrifuge, the rotating speed of 5000 rev/min is centrifuged within second day
15min, absorption supernatant is put into standby in the centrifuge tube that 1.5mL sterilized；
(3) adopt the serum VTG assay kit measurement VTG of cayman chemical company in 30 saddles
With the content in lithosporic fish serum, concrete continuous mode is as follows：
A () dilutes Dilution Buffer：15mL 5 × Dilution Buffer is taken to add 60mLddH2O is standby；
Wash Buffer：1 bottle of Wash Buffer is taken to add 1000mL ddH2O is standby；Standard：Take 5 μ g Standard powder
It is dissolved in 1mLddH2O, then add 4950 μ L to dilute with the Standard that liquid-transfering gun draws 50 μ L previous step dissolvings
Dilution Buffer, then half-and-half dilutes and becomes 0.05ng/mL until concentration；All dilution are all in the conjunction sterilizing
Carry out in the centrifuge tube of suitable specification, above diluent materials are all that test kit carries.
(b) dilute sample：Take 10 μ L blood serum samples to add 490 μ LDilution Buffer, then take 10 μ L previous step dilutions
Good blood serum sample adds 990 μ L Dilution Buffer, then takes the blood serum sample that 10 μ L previous steps have diluted to add 990 μ l
Dilution Buffer, so dilutes three concentration；
C () is loaded：96 orifice plates taking out in test kit have planned blank well, standard sample wells, sample well, plus 100 μ
LDilution Buffer is in blank well, plus the Standard that 100 μ L have diluted is in standard sample wells, plus 100 μ L dilute
Sample in sample well, room temperature place 1h, place 1h during dilute antibody：24 μ L antibody are added 12mL
Dilution Buffer；Washed 3 times with 300 μ L Wash Buffer after 1h, add the antibody that 100 μ L have diluted in each hole
In, room temperature is washed 5 times with 300 μ LWash Buffer after placing 1h again, adds 100 μ LTMB (test kit carries) in all holes
In, place 15 minutes in dark；Finally plus 0.3M sulphuric acid is in all holes, terminating reaction.
D () reads data, record data in microplate reader.
(4) 30 epinephelus lanceolatus fishes of 11-KT assay kit measurement of cayman chemical company are adopted
The content of 11-KT in serum, concrete grammar is as follows：
A () dilutes EIA Buffer：10mL EIA Buffer is taken to be added to 90mL ddH2In O；Wash Buffer joins
System：5mLWash Buffer mother solution is taken to be added to 1995mLddH21mL ploysorbate is added in O；EIA Trace is dilute
Release：100dtn EIA Trace is taken to add the EIA Buffer that 6mL has diluted；EIA antiserum dilutes：100dtn is taken to add
The EIA Buffer that 6mL has diluted；Standard dilutes：The Standard taking 100 μ L 10ng/mol adds 900 μ LddH2O
In, then it is taken out 100 μ L and is added in the EIA Buffer that 900 μ L have diluted, then take 500 μ L to be added to 500 μ L diluting
EIA Buffer in, then take 500 μ L of previous step to be added to the EIABuffer that 500 μ L have diluted, be diluted to successively
0.78pg/mL；Ellman Reagent dilutes：100dnt Ellman Reagent is taken to add 20mLddH2O；AchE Tracer
Dilution：100dtn AchE Tracer is taken to add the EIA Buffer that 6mL has diluted；All dilution were all sterilizing not
Carry out in the centrifuge tube of same specification, above diluent materials are all that test kit carries；
The preparation of (b) sample：Take in the sterilized centrifuge tube of 100 μ L serum 1.5mL, then draw 5 μ L serum with liquid-transfering gun
It is added to 955 μ L ddH2In O, then the serum 5 μ L of a step dilution is taken to add 955ul ddH2In O, so it is diluted to three concentration；
C () is loaded：96 orifice plates draw TA, BLK, NSB, B0, the hole of TAHE and standard substance, plus 100 μ LEIA
In Buffer to NSB hole, plus EIA Buffer to the B that 50 μ L have diluted0In, add standard substance by concentration order from big to small,
Again by the sample having diluted, points of three concentration add in three parallel holes, add 50 μ L AchE Tracer to except TAHE,
Each hole outside BLK, plus 50 μ L EIA antiserum each hole in addition to TA, NSB and BLK, rooms on plate
Temperature is washed 5 times with wash buffer after placing 2h, plus 200 μ L Ellman Reagent are in each hole, plus 5 μ L AchE
Tracer, to each hole in addition to TA, covers film dark place and puts 90～120 minutes.
D () reads data, record data in microplate reader；
(5) according to vitellogenin content in serum<0.9mg/mL and 11- ketone group testosterone concentration>1ng/mL then differentiates
The standard of milter, carries out discriminating to the result measuring and draws there are 11 milters (table 1) in 30 fishes；
(6) adopt gonad withdrawal device to extract a small amount of gonadal tissue of 11 milters being judged with the inventive method, put Bonn
In family name's liquid, carry out the accuracy that paraffin-embedded tissue is cut into slices for verifying the method, correspond to every fish when extracting gonad
Labelling is seen, section display 11 milters identifying of the present invention are all correct, present invention determine that result rate of accuracy reached is to 100%.
Table 1 VTG and 11-KT content and its result of determination to 30 fishes of Daya Gulf
Embodiment 2
Step is similar to Example 1, and its difference is that choosing the place of parent fish and time is on July 15th, 2014 in Hainan
25 body weight are chosen in the epinephelus lanceolatus fish of more than 20kg in Sanya, mark, and tail vein has extracted 4 DEG C of placements of blood of 5mL
Overnight, 4 DEG C on centrifuge, the rotating speed centrifugation 12min of 3500 rev/min obtains serum, uses kit measurement vitellogenin
With the content of 11- ketone group testosterone, the result of mensure draws there are 6 milters (table 2) in 25 fishes.Judge to be reflected by extracting gonad
6 milters not gone out are all correct.
Table 2 VTG and 11-KT content and its result of determination to 25 fishes in Sanya
Table 3 the inventive method differentiates the contrast of epinephelus lanceolatus fish milter method with conventional
To sum up, show that the inventive method has higher accuracy and larger use valency by the result of observation sample
Value, synthetic determination rate of accuracy reached to 100%, the discrimination method of the epinephelus lanceolatus fish milter that the present invention provides is compared with previous methods
There is efficiently and accurately, simple, the advantages of little to parent fish injury (table 3).This technology will have significantly in terms of fish breeding
Economic benefit and social benefit.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention are not subject to above-described embodiment
Limit.Other any spirit without departing from the present invention and the change made under principle, modification, replacement, combine, simplify,
All should be equivalent substitute mode, be included in protection scope of the present invention.
Claims (5)
1. a kind of method differentiating male epinephelus lanceolatus fish, is characterized in that comprising the following steps：
(1) select epinephelus lanceolatus fish：The epinephelus lanceolatus fish in selected reproduction season；
(2) obtain epinephelus lanceolatus fish serum：Choose epinephelus lanceolatus fish tail venous blood, after centrifugation, obtain serum；
(3) measure vitellogenin content in serum：Using the vitellogenin content in enzyme-linked immunosorbent assay serum：
(4) measure 11- ketone group testosterone concentration in serum：Contained using the 11- ketone group testosterone in enzyme-linked immunosorbent assay serum
Amount；
(5) differentiate epinephelus lanceolatus fish milter：Contain in conjunction with vitellogenin content in epinephelus lanceolatus fish serum and 11- ketone group testosterone
Amount, identifies epinephelus lanceolatus fish milter；
As vitellogenin content ＜ 0.9mg/mL and 11- ketone group testosterone concentration ＞ in epinephelus lanceolatus fish serum in step (5)
During 1ng/mL, differentiate as epinephelus lanceolatus fish milter.
2. the method differentiating male epinephelus lanceolatus fish according to claim 1, is characterized in that：Body weight is selected in step (1)
In the epinephelus lanceolatus fish of the mating period of more than 20kg, mating period south coastal for 5 annual～August.
3. the method differentiating male epinephelus lanceolatus fish according to claim 1, is characterized in that：In step (2) during centrifugation from
Scheming rotating speed is 3500～5000rpm, and centrifuging temperature is 4 DEG C, and centrifugation time is 10～15min.
4. the method differentiating male epinephelus lanceolatus fish according to claim 1, is characterized in that：Enzyme is adopted to join in step (3)
When immunoabsorption measures the vitellogenin content in serum, using the serum yolk of cayman chemical company
Proteinogen assay kit measurement.
5. the method differentiating male epinephelus lanceolatus fish according to claim 1, is characterized in that：Enzyme is adopted to join in step (4)
When immunoabsorption measures the 11- ketone group testosterone concentration in serum, using the serum 11- of cayman chemical company
Ketone group testosterone concentration measures kit measurement.
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| CN111018967B (en) * | 2019-12-31 | 2020-09-22 | 广东越群海洋生物研究开发有限公司 | Epinephelus lanceolatus insulin-like factor INSL-3 gene, encoding protein and application thereof |
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| CN102707073A (en) * | 2012-06-21 | 2012-10-03 | 江苏大学 | Enzyme linked immunosorbent assay kit for detecting metandienone and using method of enzyme linked immunosorbent assay kit |
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| CN104808005A (en) | 2015-07-29 |
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