CN108680750A - The ELISA detection method and kit of TROP2 expressing quantities - Google Patents
The ELISA detection method and kit of TROP2 expressing quantities Download PDFInfo
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Abstract
The present invention relates to biomedicine technical field, the specifically ELISA detection method and kit of TROP2 expressing quantities, the ELISA kit of TROP2 expressing quantities includes that TROP2 antibody is coated with 96 hole reaction plates；TROP2 antibody HRP enzyme-labeled secondary antibodies；TMB developing solutions；H2SO4 terminate liquids.The ELISA detection method and kit of a kind of TROP2 expressing quantities provided by the invention, the detection method can quickly detect TROP2 expressing quantities in serum, the high expression in various Serum of Cancer Patients that researches show that CD151 albumen.It prompts TROP2 albumen as a kind of potential serodiagnosis marker, can judge for clinical quick diagnosis, therapeutic effect and Index for diagnosis provides reference.
Description
Technical field
The present invention relates to biomedicine technical field, the ELISA detection method of specifically a kind of TROP2 expressing quantities and
Kit.
Background technology
Malignant tumour has become the principal disease for seriously threatening human health.China's tumour occurs number and constantly increases,
The death rate rises to the 108.26/10 ten thousand of the nineties from the 83.65/10 ten thousand of the seventies, rises 29.42%.Guangxi it is pernicious
Tumor mortality rate is also in rising trend, and (34700 people are dead) just increased by 12700 people than 1991 within only 1996, increased close
57.73%；The mortality of malignant tumors mark YPLL rate in Guangxi is 9.88 ‰；Residents in Guangxi all one's life (being calculated to 74 years old by birth) is dead
In the risk of malignant tumour be about 1/10 or so (cumulative risk 10.66%)；Malignant tumour makes the Guangxi population average expectation longevity
Life reduces 2.13 years old.Malignant tumour has caused heavy social economical burden, it is estimated that being used for the payment for medical care of cancer patient every year
With about 80,000,000,000 yuan, account for about the 20% of Health Expenditure.The whole nation is 185.1 ten thousand man-years because the disability that cancer is lost adjusts life years,
It is up to 1432.3 hundred million yuan with the economic loss that this is estimated.The early diagnosis of tumour early control be current prophylactic treatment tumour effective means.
TROP2 is epithelial cell membrane surface glycoprotein receptor, and one of function is the cross-film turn as intracellular calcium signal
Guide.TROP2 is very low without expression or expression in the normal tissue, and is then overexpressed in various cancerous tissues, therefore is swollen
The significantly marker of tumor diagnosis, treatment and one of prognostic evaluation.In recent years the study found that the expression of TROP2 with
Carcinoma of endometrium, oophoroma, prostate cancer, cancer of pancreas, colorectal cancer, stomach and the cancer of the esophagus, cholangiocarcinoma, carcinoma of mouth, glioma
Invasion and transfer with breast cancer have substantial connection.TROP2 albumen is in addition to other than endoglin expression, the extracellular segment portion of molecule
Dividing can occur to hydrolyze and be discharged into blood circulation under the action of enzyme, and may react the state of development of tumour, be a kind of potential
Serodiagnosis marker.Expression and the serological index detection for carrying out TROP2 albumen, are conducive to carry out clinical quick
Diagnosis, therapeutic effect judge and the research and application of Index for diagnosis.Currently, the TROP2 detection methods reported both at home and abroad are all not bery
Maturation is simply used for scientific experiment, not yet develops into the kit suitable for clinical detection, is unfavorable for routine clinical use.
Invention content
It is an object of the invention in view of the above-mentioned drawbacks in the prior art, provide a kind of TROP2 expressing quantities
ELISA detection methods and kit.
For achieving the above object, present invention employs following technical solutions：
A kind of ELISA kit of TROP2 expressing quantities, including TROP2 antibody are coated with 96 hole reaction plates；TROP2 is anti-
Body-HRP enzyme-labeled secondary antibodies；TMB developing solutions；H2SO4Terminate liquid；Cleaning solution；ELISA kit forms：
In addition, the present invention also provides following attached technical schemes：
Preferably, the ELISA detection method of the TROP2 expressing quantities, includes the following steps：
(1) ELISA reaction reagents and optimization reaction condition are prepared：Prepare monoclonal antibody hybridoma-mouse ascites model；It is single
The purifying of anti-hybridoma-mouse ascites TROP2 antibody；TROP2 antibody-HRP enzyme label screenings；Purify TROP2 antibody-HRP enzymes
Mark conjugate；
(2) purifying of monoclonal antibody hybridoma-mouse ascites TROP2 antibody；TROP2 antibody-HRP enzyme label screenings；Purifying
TROP2 antibody-HRP enzymes mark conjugate；
(3) TROP2 albumen in normal control and sample to be tested serum is detected using elisa technique；
(4) antigen-antibody stays after combining in solid phase carrier；
(5) product is formed after being reacted with enzyme labelled antibody；
(6) after developing solution reaction is added, H is added2SO4Reaction is terminated, interpretation of result is carried out according to the depth of color products.
Preferably, the ELISA detection method of the TROP2 expressing quantities, using elisa technique to normal serum or
Sample serum to be checked is detected, and specific method is：
A, normal serum or sample serum to be checked are taken, first time incubation is carried out with the TROP2 antibody of surface of solid phase carriers, uses
The method of washing makes the antigen-antibody complex formed on solid phase carrier be separated with other substances in liquid；
B, HRP enzymic-labelled antibodies are added and carry out second of incubation.
Preferably, TROP2 antibody is diluted to 2 μ g/mL in step A, wrapper sheet 100uL, wrapper sheet condition be 2~5 DEG C place 8~
15 hours.
Preferably, 100uL serum to be checked is taken to be added in orifice plate in step A, PBST cleanings 3~5 times after 40~50min, often
It has the final say after secondary board-washing.
Preferably, TROP2 antibody first time incubation temperature is 37 DEG C in step A, and the time is 40~50min, is being used
PBST is cleaned 3~5 times, is had the final say after each board-washing.
Preferably, second of incubation temperature of TROP2 antibody is 37 DEG C in step B, and the time is 40~50min, is being used
PBST is cleaned 3~5 times, is had the final say after each board-washing.
Preferably, the ELISA detection method of the TROP2 expressing quantities, after cleaning be added 90~
40~60uL2M H2SO4 are added after 4~8min and terminate reaction, obtain experimental result for 110uLTMB developing solutions.
TROP2 albumen is very low without expression or expression in the normal tissue, and is then overexpressed in various cancerous tissues.Its
The extracellular fragmented portion of molecule can occur to hydrolyze and be discharged into blood circulation under the action of enzyme, and may reflect the development of tumour
State is a kind of potential serodiagnosis and monitoring marker.
Advantageous effect of the present invention is：A kind of ELISA detection method of TROP2 expressing quantities provided by the invention and examination
Agent box, the detection method can quickly detect TROP2 expressing quantities in serum, and researches show that TROP2 albumen in various tumours
High expression in patients serum.It prompts TROP2 albumen as a kind of potential serodiagnosis marker, can be that clinic is quickly examined
Disconnected, therapeutic effect judges and Index for diagnosis provides reference.
Description of the drawings
Fig. 1 is the ELISA results of TROP2 standard antigens；
Fig. 2 is the sensitivity Detection of TROP2 double antibody sandwich methods；
Fig. 3 A are the expression of TROP2 in blood serum of patients with human breast carcinoma sample；
Fig. 3 B are that TROP2 is expressed in normal health control serum sample.
Specific implementation mode
Technical solution of the present invention is further non-limitingly described in detail below in conjunction with preferred embodiment.
Embodiment 1
The ELISA kit of TROP2 expressing quantities, including TROP2 antibody are coated with 96 hole reaction plates；TROP2 antibody-
HRP enzyme-labeled secondary antibodies；TMB developing solutions；H2SO4 terminate liquids；Cleaning solution；ELISA kit forms：
The ELISA detection method of the TROP2 expressing quantities, includes the following steps：
(1) ELISA reaction reagents and optimization reaction condition are prepared：Prepare monoclonal antibody hybridoma-mouse ascites model；It is single
The purifying of anti-hybridoma-mouse ascites TROP2 antibody；TROP2 antibody-HRP enzyme label screenings；Purify TROP2 antibody-HRP enzymes
Mark conjugate；
(2) purifying of monoclonal antibody hybridoma-mouse ascites TROP2 antibody；TROP2 antibody-HRP enzyme label screenings；Purifying
TROP2 antibody-HRP enzymes mark conjugate；
(3) TROP2 albumen in normal control and sample to be tested serum is detected using elisa technique；
(4) antigen-antibody stays after combining in solid phase carrier；
(5) product is formed after being reacted with enzyme labelled antibody；
(6) after developing solution reaction is added, H2SO4 is added and terminates reaction, interpretation of result is carried out according to the depth of color products.
The ELISA detection method of the TROP2 expressing quantities, using elisa technique to normal serum or mark to be checked
This serum is detected, and specific method is：
A, normal serum or sample serum to be checked are taken, first time incubation is carried out with the TROP2 antibody of surface of solid phase carriers, uses
The method of washing makes the antigen-antibody complex formed on solid phase carrier be separated with other substances in liquid；
B, HRP enzymic-labelled antibodies are added and carry out second of incubation.
TROP2 antibody is diluted to 2 μ g/mL, wrapper sheet 100uL in step A, and wrapper sheet condition is 2 DEG C and places 8 hours.
100uL serum to be checked is taken to be added in orifice plate in step A, PBST is cleaned 3 times after 40min, is had the final say after each board-washing.
TROP2 antibody first time incubation temperature is 37 DEG C, time 45min in step A, is using PBST cleanings 4 times, often
It has the final say after secondary board-washing.
Second of incubation temperature of TROP2 antibody is 37 DEG C, time 40min in step B, is using PBST cleanings 4 times, often
It has the final say after secondary board-washing.
110uLTMB developing solutions are added in the ELISA detection method of the TROP2 expressing quantities after cleaning,
40uL2M H2SO4 are added after 4min and terminate reaction, obtain experimental result.
Embodiment 2
The ELISA kit of TROP2 expressing quantities, including TROP2 antibody are coated with 96 hole reaction plates；TROP2 antibody-
HRP enzyme-labeled secondary antibodies；TMB developing solutions；H2SO4 terminate liquids；ELISA kit forms：
The ELISA detection method of the TROP2 expressing quantities, includes the following steps：
(1) ELISA reaction reagents and optimization reaction condition are prepared：Prepare monoclonal antibody hybridoma-mouse ascites model；It is single
The purifying of anti-hybridoma-mouse ascites TROP2 antibody；TROP2 antibody-HRP enzyme label screenings；Purify TROP2 antibody-HRP enzymes
Mark conjugate；
(2) purifying of monoclonal antibody hybridoma-mouse ascites TROP2 antibody；TROP2 antibody-HRP enzyme label screenings；Purifying
TROP2 antibody-HRP enzymes mark conjugate；
(3) TROP2 albumen in normal control and sample to be tested serum is detected using elisa technique；
(4) antigen-antibody stays after combining in solid phase carrier；
(5) product is formed after being reacted with enzyme labelled antibody；
(6) after developing solution reaction is added, H2SO4 is added and terminates reaction, interpretation of result is carried out according to the depth of color products.
The ELISA detection method of the TROP2 expressing quantities, using elisa technique to normal serum or mark to be checked
This serum is detected, and specific method is：
A, normal serum or sample serum to be checked are taken, first time incubation is carried out with the TROP2 antibody of surface of solid phase carriers, uses
The method of washing makes the antigen-antibody complex formed on solid phase carrier be separated with other substances in liquid；
B, HRP enzymic-labelled antibodies are added and carry out second of incubation.
TROP2 antibody is diluted to 2 μ g/mL, wrapper sheet 100uL in step A, and wrapper sheet condition is 3 DEG C and places 10 hours.
100uL serum to be checked is taken to be added in orifice plate in step A, PBST is cleaned 5 times after 50min, is had the final say after each board-washing.
TROP2 antibody first time incubation temperature is 37 DEG C, time 50min in step A, is using PBST cleanings 5 times, often
It has the final say after secondary board-washing.
Second of incubation temperature of TROP2 antibody is 37 DEG C, time 50min in step B, is using PBST cleanings 5 times, often
It has the final say after secondary board-washing.
100uLTMB developing solutions are added in the ELISA detection method of the TROP2 expressing quantities after cleaning,
60uL2M H2SO4 are added after 5min and terminate reaction, obtain experimental result.
Embodiment 3
The ELISA kit of TROP2 expressing quantities, including TROP2 antibody are coated with 96 hole reaction plates；TROP2 antibody-
HRP enzyme-labeled secondary antibodies；TMB developing solutions；H2SO4 terminate liquids；Cleaning solution；ELISA kit forms：
The ELISA detection method of the TROP2 expressing quantities, includes the following steps：
(1) ELISA reaction reagents and optimization reaction condition are prepared：Prepare monoclonal antibody hybridoma-mouse ascites model；It is single
The purifying of anti-hybridoma-mouse ascites TROP2 antibody；TROP2 antibody-HRP enzyme label screenings；Purify TROP2 antibody-HRP enzymes
Mark conjugate；
(2) purifying of monoclonal antibody hybridoma-mouse ascites TROP2 antibody；TROP2 antibody-HRP enzyme label screenings；Purifying
TROP2 antibody-HRP enzymes mark conjugate；
(3) TROP2 albumen in normal control and sample to be tested serum is detected using elisa technique；
(4) antigen-antibody stays after combining in solid phase carrier；
(5) product is formed after being reacted with enzyme labelled antibody；
(6) after developing solution reaction is added, H2SO4 is added and terminates reaction, interpretation of result is carried out according to the depth of color products.
The ELISA detection method of the TROP2 expressing quantities, using elisa technique to normal serum or mark to be checked
This serum is detected, and specific method is：
A, normal serum or sample serum to be checked are taken, first time incubation is carried out with the TROP2 antibody of surface of solid phase carriers, uses
The method of washing makes the antigen-antibody complex formed on solid phase carrier be separated with other substances in liquid；
B, HRP enzymic-labelled antibodies are added and carry out second of incubation.
TROP2 antibody is diluted to 2 μ g/mL, wrapper sheet 100uL in step A, and wrapper sheet condition is 5 DEG C and places 10 hours.
100uL serum to be checked is taken to be added in orifice plate in step A, PBST is cleaned 4 times after 45min, is had the final say after each board-washing.
TROP2 antibody first time incubation temperature is 37 DEG C, time 45min in step A, is using PBST cleanings 4 times, often
It has the final say after secondary board-washing.
Second of incubation temperature of TROP2 antibody is 37 DEG C, time 45min in step B, is using PBST cleanings 4 times, often
It has the final say after secondary board-washing.
110uLTMB developing solutions are added in the ELISA detection method of the TROP2 expressing quantities after cleaning,
60uL2M H2SO4 are added after 8min and terminate reaction, obtain experimental result.
Embodiment 4
The ELISA kit of TROP2 expressing quantities, including TROP2 antibody are coated with 96 hole reaction plates；TROP2 antibody-
HRP enzyme-labeled secondary antibodies；TMB developing solutions；H2SO4 terminate liquids；ELISA kit forms：
。
The ELISA detection method of the TROP2 expressing quantities, includes the following steps：
(1) ELISA reaction reagents and optimization reaction condition are prepared：Prepare monoclonal antibody hybridoma-mouse ascites model；It is single
The purifying of anti-hybridoma-mouse ascites TROP2 antibody；TROP2 antibody-HRP enzyme label screenings；Purify TROP2 antibody-HRP enzymes
Mark conjugate；
(2) purifying of monoclonal antibody hybridoma-mouse ascites TROP2 antibody；TROP2 antibody-HRP enzyme label screenings；Purifying
TROP2 antibody-HRP enzymes mark conjugate；
(3) TROP2 albumen in normal control and sample to be tested serum is detected using elisa technique；
(4) antigen-antibody stays after combining in solid phase carrier；
(5) product is formed after being reacted with enzyme labelled antibody；
(6) after developing solution reaction is added, H2SO4 is added and terminates reaction, interpretation of result is carried out according to the depth of color products.
The ELISA detection method of the TROP2 expressing quantities, using elisa technique to normal serum or mark to be checked
This serum is detected, and specific method is：
A, normal serum or sample serum to be checked are taken, first time incubation is carried out with the TROP2 antibody of surface of solid phase carriers, uses
The method of washing makes the antigen-antibody complex formed on solid phase carrier be separated with other substances in liquid；
B, HRP enzymic-labelled antibodies are added and carry out second of incubation.
TROP2 antibody is diluted to 2 μ g/mL, wrapper sheet 100uL in step A, and wrapper sheet condition is 2 DEG C and places 8 hours.
100uL serum to be checked is taken to be added in orifice plate in step A, PBST is cleaned 4 times after 45min, is had the final say after each board-washing.
TROP2 antibody first time incubation temperature is 37 DEG C, time 45min in step A, is using PBST cleanings 4 times, often
It has the final say after secondary board-washing.
Second of incubation temperature of TROP2 antibody is 37 DEG C, time 45min in step B, is using PBST cleanings 4 times, often
It has the final say after secondary board-washing.
90~110uLTMB colour developings are added in the ELISA detection method of the TROP2 expressing quantities after cleaning
40~60uL2M H2SO4 are added after 4~8min and terminate reaction, obtain experimental result for liquid.
Test example 1
Refer to that the serum color reaction to be checked is higher than blank control and negative control, then in the positive of the present invention
TROP2 protein expressions are positive.
Refer to that the serum color reaction to be checked is right higher than blank control and feminine gender in expression quantity height of the present invention
According to, and its color reaction is deeper, TROP2 expressing quantities are higher.
Biomaterial used, examination in the ELISA detection method and kit of TROP2 expressing quantities provided by the invention
Agent or instrument are available on the market.
One, material and method
1, patients serum's sample
In order to carry out EILSA detections, The People's Hospital, Guangxi Zhuang Autonomous Region in December, -2016 in January, 2014 is collected altogether
The serum specimen of 46 patient with breast cancers and 46 normal health physical examinations control.Collection standard is the pathological diagnosis of breast cancer,
First visit and untreated, without the medical history of other tumours.Carcinoma in situ excludes except research.Clinical sample is only used for studying,
Ratify by Ethics Committee of The People's Hospital, Guangxi Zhuang Autonomous Region.
Clinical sample include 46 Female breast cancer patients, the age between 35-55 Sui, average age 42.6 years old.It is all
Patient receives mammary gland carcinectomy.The histopathology histological grading and clinical stages of this research according to the World Health Organization (WHO) and
The standard of sixth version pTMN International Union Against Cancers (UICC, 2002) defines.By two diseases of The People's Hospital, Guangxi Zhuang Autonomous Region
Reason doctor assesses the histological type of all tumours and classification.
2、ELISA
By reagent 1：96 hole enzyme labelled antibody coating plates take out from 4 DEG C, place 5-10min at room temperature；By reagent 2：
The TROP2 antigen standards for knowing concentration, are diluted, concentration is respectively after dilution with PBS：0,3.125,6.25,12.5,25,
50,100ng/ml.By the TROP2 antigens of known concentration, 10ug/ml dilutes 1 with 1 × PBS:100.Each ELISA plates use
5ul carries out 1:100 dilutions, the first final concentration of 100ng/ml of pipe, then press 1：2 serial dilutions keep last pipe final concentration of
3.125ng/ml.Each 100uL of TROP2 antigens and sample to be tested of known concentration is added in ELISA microwell plates, incubation at room temperature
Time is 45min；By reagent 3：10 × PBS and reagent 4：Tween-20 takes out from refrigerator, the standard of PBST in preparing by reagent
Prepared by Preparation Method, a plate probably needs 360ml, PBST；ELISA microwell plates are cleaned with PBST 4 times, per hole 300ul, every time
It has the final say after board-washing；After cleaning, reagent 5 is added：No. TROP2-37 clone's enzyme labelled antibody, per hole 100ul.Enzyme labelled antibody
It is diluted with 1XPBS, exact dilution multiple is 1/40 (final concentration 2ug/ml), and extension rate is improper to will produce background, incubation at room temperature
45min；It is cleaned 4 times with PBST, is had the final say after each board-washing；After cleaning, reagent 6 is added：TMB developing solutions are shown
Color is incubated at room temperature 5min per hole 100ul；It is accurately incubated 3.5min at room temperature, reagent 7 is added：2M H2SO4Reaction is terminated, per hole
50ul, incubation time are more than that 3.5min will produce background；With microplate reader in OD450nmMeasure the OD in each hole450nmValue.
3, recruitment evaluation
TROP2 shows good linear relationship within the scope of a certain concentration, will present out " S " curve beyond the range.Cause
This shows as OD values and is higher than highest point, then need pair if the antigen concentration of TROP2 is higher than above range in sample segment
ELISA reactions are carried out after sample dilution.Typical result is illustrated in fig. 1 shown below：Various concentration (0,3.125,6.25,12.5,25,
50,100ng/ml, sequence is from left to right)
4, statistical analysis
TROP2 expressing quantities in assessment breast cancer patients and normal health physical examination control serum are examined with t.Use SPSS
22.0 softwares carry out statistical analysis.P ＜ 0.05, it is believed that difference is significant.
Two, result
1, breast cancer patients TROP2 high is expressed
It is differential expressions of the analysis TROP2 between patient with breast cancer and normal health compare, this research ELISA method
Detect the expression of TROP2 in the serum sample of 46 patient with breast cancers and the control of 46 normal healths.Wherein 25
(54.3%) TROP2 expresses (Fig. 3 A) in high in blood serum of patients with human breast carcinoma sample；In 6 normal health control serum samples
TROP2 expresses (6.5%) (Fig. 3 B) in low expression (13.0%) and 3 in moderate strength, (P ＜ 0.05).
OD values shown in Fig. 3 A and 3B through t inspection compares, TROP2 albumen expressed in patient with breast cancer and normal person have it is aobvious
Write sex differernce (P ＜ 0.05).
It is pointed out that the technical concepts and features of above-mentioned preferred embodiment only to illustrate the invention, its object is to
Those skilled in the art can understand the contents of the present invention and implements according to this, and the protection of the present invention can not be limited with this
Range.Any equivalent change or modification in accordance with the spirit of the invention should be covered by the protection scope of the present invention.
Claims (8)
- The ELISA kit of 1.TROP2 expressing quantities, it is characterised in that：It is coated with 96 hole reaction plates including TROP2 antibody； TROP2 antibody-HRP enzyme-labeled secondary antibodies；TMB developing solutions；H2SO4 terminate liquids；Cleaning solution；ELISA kit forms：。
- 2. the ELISA detection method of TROP2 expressing quantities according to claim 1, it is characterised in that：Including walking as follows Suddenly：（1）Prepare ELISA reaction reagents and optimization reaction condition：Prepare monoclonal antibody hybridoma-mouse ascites model；Monoclonal antibody is miscellaneous Hand over the purifying of tumor-mouse ascites TROP2 antibody；TROP2 antibody-HRP enzyme label screenings；Purify TROP2 antibody-HRP enzymes label Conjugate；（2）The purifying of monoclonal antibody hybridoma-mouse ascites TROP2 antibody；TROP2 antibody-HRP enzyme label screenings；It is anti-to purify TROP2 Body-HRP enzymes mark conjugate；（3）TROP2 albumen in normal control and sample to be tested serum is detected using elisa technique；（4）Antigen-antibody stays after combining in solid phase carrier；（5）Product is formed after being reacted with enzyme labelled antibody；（6）After developing solution reaction is added, H is added2SO4Reaction is terminated, interpretation of result is carried out according to the depth of color products.
- 3. the ELISA detection method of TROP2 expressing quantities according to claim 1, it is characterised in that：The use Elisa technique is detected normal serum or sample serum to be checked, and specific method is：A, normal serum or sample serum to be checked are taken, first time incubation is carried out with the TROP2 antibody of surface of solid phase carriers, with washing Method so that the antigen-antibody complex formed on solid phase carrier is separated with other substances in liquid；B, HRP enzymic-labelled antibodies are added and carry out second of incubation.
- 4. the ELISA detection method of TROP2 expressing quantities according to claim 3, it is characterised in that：In step A TROP2 antibody is diluted to 2 μ g/mL, wrapper sheet 100uL, and wrapper sheet condition is 2 ~ 5 DEG C and places 8 ~ 15 hours.
- 5. the ELISA detection method of TROP2 expressing quantities according to claim 4, it is characterised in that：It is taken in step A 100uL serum to be checked is added in orifice plate, and PBST is cleaned 3 ~ 5 times after 40 ~ 50min, has the final say after each board-washing.
- 6. the ELISA detection method of TROP2 expressing quantities according to claim 3, it is characterised in that：In step A TROP2 antibody first time incubation temperatures are 37 DEG C, and the time is 40 ~ 50min, is cleaned 3 ~ 5 times using PBST, is clapped after each board-washing Plate.
- 7. the ELISA detection method of TROP2 expressing quantities according to claim 3, it is characterised in that：In step B Second of incubation temperature of TROP2 antibody is 37 DEG C, and the time is 40 ~ 50min, is cleaned 3 ~ 5 times using PBST, is clapped after each board-washing Plate.
- 8. according to the ELISA detection method of claim 5 ~ 7 any one of them TROP2 expressing quantities, it is characterised in that： 90 ~ 110uLTMB developing solutions are added after cleaning, 40 ~ 60uL2M H2SO4, which are added, after 4 ~ 8min terminates reaction, is tested As a result.
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| CN112180101A (en) * | 2020-06-01 | 2021-01-05 | 北京松果天目健康管理有限公司 | Kit for detecting abundance of ACE2 receptor and detection method |
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