CN107677822A - Detect the fluorecyte count detection kit of tumor markers - Google Patents
Detect the fluorecyte count detection kit of tumor markers Download PDFInfo
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- CN107677822A CN107677822A CN201610633992.0A CN201610633992A CN107677822A CN 107677822 A CN107677822 A CN 107677822A CN 201610633992 A CN201610633992 A CN 201610633992A CN 107677822 A CN107677822 A CN 107677822A
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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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
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Abstract
The present invention relates to field of biological medicine, specifically, it is related to a kind of fluorecyte count detection kit for detecting tumor markers, including to mark the CD14 antibody of Cell membrane antigens and CD16 antibody, and to mark the antibody of endochylema internal antigens；The endochylema internal antigens are TKTL1 and/or DNASE1L1；The kit also includes microslide.
Description
Technical field
The present invention relates to field of biological medicine, in particular to a kind of fluorecyte for detecting tumor markers
Count detection kit.
Background technology
Monocytes/macrophages include huge in the premonocyte in marrow, the monocyte in peripheral blood and tissue
Phagocyte.Human peripheral's haemocyte is broadly divided into cytode (predominantly erythrocyte) and karyocyte (leucocyte etc.).It is single
Nucleus accounts for 5% or so of karyocyte sum, is antigen submission in immune system by medullary system hematopoietic progenitor cell
The important component of cell (APC cells).
In human peripheral blood circulation, leucocyte is distributed in two places, i.e. circulatory pool and storage pool, circulatory pool is exactly
Leucocyte in fluid flow blood, storage pool are just attached to the leucocyte of vascular wall, lymph node, marrow etc., are temporarily not involved in
Blood circulation, but the leucocyte in the leucocyte and circulatory pool in storage pool keeps dynamic equilibrium.Leucocyte is in human circulation system
It is ripe in system to break up and realize that immunologic function be roughly divided into following steps：Monocyte in capillary → pass through blood
Tissue outside tube wall intravasation → discovery heterologous material (bacterium, pathogen, tumour cell etc.) → is divided into the macrophage of maturation
Cell → phagocytosis and digest → by antigen submission to specific cell surface molecule → via lymphatic vessel be back to peripheral blood →
Complete antigen submission and activate lethal immunocyte.
As the external pathogen invasion of fight and the sentry for understanding itself abnormal cell mechanism, macrophage is immune
System forms the important step of immune response, and effect is great.Have all the time in normal human towards anti-tumor (immortality
Change, improper apoptosis) cell of development produces, and normal immunity of organism mechanism can be removed it in time by said process.
As can be seen here no matter phagocytosis of the macrophage for anti-tumor cell in healthy population or cancer patient's body, when being all no
Without carving not in progress, for the observation of tumour source material in monocytes/macrophages (including protein, accounting, lipid etc.),
The new perspective that a monitoring in-vivo tumour occurs, developed can be provided for us, its application prospect is necessarily very extensive.
Important component of the monocytes/macrophages as karyocyte, laboratory is generally by its surface antigen molecules
What CD14 and CD16 was detected, it is currently available the antibody of maturation.Main clinical practice concentrates on haemocyte composition inspection
Survey with hematopoietic system cancer (such as：Grain/monocytic leukemia) in terms of detection in, provide important references for clinical diagnosis.
But it is associated with the monitoring of entity tumor, from the point of view of current patent and literature search, still lacks enough systematicness
Correlative study, and not yet form matured product, also there is no Patents mandate in CONTINENTAL AREA OF CHINA.
In view of this, it is special to propose the present invention.
The content of the invention
It is an object of the invention to provide a kind of fluorecyte count detection kit for detecting tumor markers, to solve
Above mentioned problem.
In order to realize the above-mentioned purpose of the present invention, spy uses following technical scheme：
A kind of fluorecyte count detection kit for detecting tumor markers, including to mark Cell membrane antigens
CD14 antibody and CD16 antibody, and to mark the antibody of endochylema internal antigens；
The endochylema internal antigens are TKTL1 and/or DNASE1L1；
The kit also includes microslide.
The principle of the present invention is the CD14 gone out with CD14 and CD16 antibody labeling in object peripheral blood to be detected+CD16+
Cell, (resisted after removing most of impurity in addition to leucocyte with antitumor mark X to be detected inside i.e. above-mentioned endochylema
It is former) the antibody labeling tumor markers；
Fluorecyte calculating instrument detects X+CD14+CD16+Cell and CD14+CD16+Cell quantity and count the former with after
The ratio of person's cell quantity.
Fluorecyte calculating instrument detection method can quickly and accurately identify and count the cell among cell mass.Inserting
After slide, fluorecyte calculating instrument can be focused and be counted to cell automatically.
By using fluorecyte calculating instrument to CD14+CD16+And X+CD14+CD16+The quantity of positive cell is detected,
It can quickly, precision evaluate global tumor markers expression.
Have determined the monocyte that different subtype in human peripheral blood be present at present.It is known mainly according to CD14 and
The expression of CD16 antigens is divided into 4 hypotypes：CD14++CD16-(G1 hypotypes), CD14++CD16+(G2 hypotypes), CD14+CD16-(G3
Hypotype), CD14+CD16+(G4 hypotypes).CD14+CD16+It is one of wherein most important hypotype, it is thinner than the monokaryon of other hypotypes
Born of the same parents have stronger phagocytic activity, thus for the principle angle of the present invention, detect CD14+CD16+The monocyte of hypotype is more
It is significant.
Tumor markers (TM), refer to that characteristic is present in malignant cell, or produced extremely by malignant cell
Material, or host to the stimulate the reaction of tumour caused material, and tumorigenesis can be reflected, monitor tumour pair
A kind of material of therapeutic response.Work as CD14+CD16+When cell swallows tumour cell, tumor markers will be in CD14+CD16+Carefully
Born of the same parents are enriched with, and CD14+CD16+Cell is the shuttling movement in human body, thus can be by detecting CD14+CD16+Tumour in cell
Mark content carrys out the convenient Tumor Marker Levels characterized in human body.
The difference of tumour cell and normal somatic cell, it is main at two aspects：
Metabolism：Tumour cell is often shown to energy because its propagation is fast and is not regulated and controled by normal cell-cycle
The demand of the formula of hungering and thirst of quantity of material (glucose etc.), but due to the variation of its metabolic mechanism, metabolism particularly energetic supersession
Efficiency is often far below normal somatic cell, is utilized for example, fully effective metabolism can not be carried out to glucose, but a kind of waste
The consumption of formula.And enzyme material of the tumour cell required in this metabolic process, its expression quantity and normal somatic cell there is also
Huge difference, the break-through point that can be detected as tumour cell.
Apoptosis：Another feature that tumour cell is different from normal somatic cell is exactly to immortalize, and tumour cell is not
By normal cell-cycle regulation and control.Apoptosis Mechanism, which has had, more clearly to be studied, tumour cell not apoptosis, that certainty
Cause the expression quantity for the factor, semiochemicals or the enzyme that regulating cell apoptosis either plays a key effect in apoptosis process
There is obvious exception, this can also serve as the break-through point of tumour cell detection.
Two tumor markerses preferred for this invention are related to metabolism with Apoptosis respectively.
TKTL1 (transketolase 1) is transketolase 1, it, the non-oxide way in main regulation glycometabolism approach
The developing stage of footpath, the high expression in kinds of tumors, its expression quantity and tumour, invasion and attack and transfer, and prognosis have substantial connection.
Nutrition arrangement and follow-up treatment to tumor patient etc. have important suggesting effect.
DNASE1L1 (Deoxyribonuclease I-Linke 1) plays an important role in apoptosis process, but
In tumour cell, normal Apoptosis mechanism failure, DNASE1L1 activity is suppressed, but its great expression and in tumour cell
Accumulation.Its expression quantity can be as the index for differentiating tumour cell or Apoptosis mechanism abnormal cell.
Preferably, fluorecyte count detection kit as described above, the fluorecyte count detection kit is also
Including fluorescence secondary antibody corresponding to one or more antibody in the CD14 antibody, CD16 antibody and internal antigens antibody.
It is further preferred that the fluorescence secondary antibody is the antibody of F (ab') 2.
The present invention needs to mark CD14+CD16+TKTL1 and/or DNASE1L1 antigens inside cell cytosol, due to F
(ab') 2 antibody do not have Fc sections, and the ability of penetration cell film is stronger, and specificity is more preferable, thus the preferably antibody of F (ab') 2 is as glimmering
Light secondary antibody.
Meanwhile in order to reduce operating procedure, it is preferred that the CD14 antibody, CD16 antibody and internal antigens antibody are
Fluorescent labeled antibody, and fluorescence color is different.
Further, no matter for fluorescence secondary antibody or fluorescence primary antibody, it is preferred that the fluorescence labeling be PE, CY5,
Any of CY7, per-CP and TRITC.
Preferably, fluorecyte count detection kit as described above, the fluorecyte count detection kit is also
Including the IgG antibody with the internal antigens antibody with Species origin.
Effect with the IgG antibody of Species origin is, can mark non-specific with internal antigens antibody non-specific binding
Property material, thus can using its signal as background signal deduct, be specifically shown in the embodiment of the present invention.
Preferably, fluorecyte count detection kit as described above, the fluorecyte count detection kit is also
Including any one or more in lysate, closing and punching buffer solution, fixer, rinsing liquid.
It is further preferred that the lysate is working solution or its mother liquor containing each composition of following concentration：
8mM~12mM Tris-HCL, 8mM~12mM NaH2PO4And/or NaHPO4, 120mM~140mM NaCl, volume
Percentage is 0.7%~1.3% Triton X-100,8mM~12mM Na4P2O7·10H2O, pH=7.2~7.7.
It is further preferred that the closing and punching buffer solution are the working solution containing each composition of following concentration or it is female
Liquid：
The Triton X-100 that 0.008g/mL~0.012g/mL BSA and percentage by volume is 0.7%~1.3% are molten
Liquid, solvent are 1 × TBS or 1 × PBS solution.
It is further preferred that the fixer is working solution or its mother liquor containing each composition of following concentration：
With volume percent, the mixed solution containing 0.8%~1.2% formaldehyde and 0.33%~0.38% methanol,
Solvent for use is water.
It is further preferred that the rinsing liquid is the working solution or its mother liquor of each composition containing following concentration：
1 × PBS solution.
Compared with prior art, beneficial effects of the present invention are：
1), the invention provides detection tumour antigen kit, the kit be directed to blood in TKTL1 and
DNASE1L1 is detected, and then knows the expression of object overall situation tumour antigen to be measured, thus available for the early stage of tumour
Diagnosis.
2), fluorecyte calculating instrument detection method can quickly and accurately identify and count the cell among cell mass.Inserting
After entering slide, fluorecyte calculating instrument can be focused and be counted to cell automatically.By using fluorecyte calculating instrument
To CD14+CD16+And X+CD14+CD16+The quantity of positive cell is detected, and can quickly, precision evaluate global tumour mark
Will thing expression.
Embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the present invention.It is unreceipted specific in embodiment
Condition person, the condition suggested according to normal condition or manufacturer are carried out.Agents useful for same or the unreceipted production firm person of instrument, it is
The conventional products that can be obtained by commercially available purchase.
Embodiment 1
1st, object peripheral blood blood sample 200ul to be measured is taken, adds antibody anti-CD14-FITC (BD companies, article No.
555397) 40ul, anti-CD16-APC (BD companies, article No. 561304) 10ul, room temperature lucifuge are incubated 15min；
2nd, add 200ul fixative room temperatures lucifuge and be incubated 8min；The composition of the fixative is：Percentage by volume is
0.8% formaldehyde, 0.30% methanol that percentage by volume is；
3rd, plus lysate 3mL is into sample, and the concussion that is vortexed mixes, and lucifuge is incubated 15min, splitting erythrocyte；The cracking
Liquid contains 8mM Tris-HCL, 8mM NaH2PO4And/or NaHPO4, 140mM NaCl, percentage by volume be 1.3% Triton
X-100、8mM Na4P2O7·10H2O, pH=7.2；
4th, 700g centrifuges 6min, removes supernatant；Supernatant slowly is outwelled, residual liquid is drawn with paper, the concussion that is vortexed is resuspended thin
Born of the same parents；
5th, 80ul closings and punching buffer solution are added, thrum mixes；
The Triton X-100 that the BSA and percentage by volume that closing and the punching buffer solution is 0.012g/mL are 1.3%
Solution；Solvent is 1 × TBS solution.
6th, above-mentioned solution system is bisected into two parts, be separately added into anti-TKTL1 (rabbit polyclonal,
Sigma, article No. HPA000505) and anti-DNASE1L1 antibody (rabbit polyclonal, sigma, article No.
HPA072635), thrum mixes, and lucifuge is incubated 20min；The adding proportion of antibody is 1:1000；
7th, 2ml PBS (8g/L NaCl, 0.2g/L KCl, 1.4g/L Na are separately added into2HPO4、0.27g/L KH2PO4)
Wash, be vortexed and mix, 800g centrifugation 5min, remove supernatant, wash 2~3 times, cell is resuspended after washing；
8th, 2ul anti-rabbit-PE antibody (donkey polyclonal, abcam companies, goods are separately added into
Number ab7007), lucifuge is incubated 10min；
9th, 2ml PBS are separately added into wash, is vortexed and mixes, 800g centrifugation 5min, supernatant is removed, washes 2~3 times, be resuspended after washing
Cell；
10th, cell is resuspended with 500ul PBS, takes 30ul sample drops on slide, smear is carried out with another concave-concave piece,
Fluorecyte calculating instrument detects
Instrument title：Thermo Fisher, Countess II FL Automated Cell Counter.
Embodiment 2
1st, object peripheral blood blood sample 200ul to be measured is taken, adds antibody anti-CD14-FITC (BD companies, article No.
555397) 40ul, anti-CD16-APC (BD companies, article No. 561304) 10ul, room temperature lucifuge are incubated 15min；
2nd, add 200ul fixative room temperatures lucifuge and be incubated 6min；The composition of the fixative is：Percentage by volume is
1.2% formaldehyde, 0.38% methanol that percentage by volume is；
3rd, plus lysate 3mL is into sample, and the concussion that is vortexed mixes, and lucifuge is incubated 15min, splitting erythrocyte；The cracking
Liquid contains 12mM Tris-HCL, 12mM NaH2PO4And/or NaHPO4, 120mM NaCl, percentage by volume be 0.7%
Triton X-100、12mM Na4P2O7·10H2O, pH=7.7；
4th, 900g centrifuges 4min, removes supernatant；Supernatant slowly is outwelled, residual liquid is drawn with paper, the concussion that is vortexed is resuspended thin
Born of the same parents；
5th, 120ul closings and punching buffer solution are added, thrum mixes；
The Triton X-100 that the BSA and percentage by volume that closing and the punching buffer solution is 0.008g/mL are 0.7%
Solution；Solvent is 1 × PBS solution.
6th, above-mentioned solution system is bisected into two parts, be separately added into anti-TKTL1 (rabbit polyclonal,
Sigma, article No. HPA000505) and anti-DNASE1L1 antibody (rabbit polyclonal, sigma, article No.
HPA072635), thrum mixes, and lucifuge is incubated 15min；The adding proportion of antibody is 1:1000；
7th, 2ml PBS (8g/L NaCl, 0.2g/L KCl, 1.4g/L Na are separately added into2HPO4、0.27g/L KH2PO4)
Wash, be vortexed and mix, 800g centrifugation 5min, remove supernatant, wash 2~3 times, cell is resuspended after washing；
8th, 2ul anti-rabbit-PE antibody (donkey polyclonal, abcam companies, goods are separately added into
Number ab7007), lucifuge is incubated 10min；
9th, 2ml PBS are separately added into wash, is vortexed and mixes, 800g centrifugation 5min, supernatant is removed, washes 2~3 times, be resuspended after washing
Cell；
10th, cell is resuspended with 500ul PBS, takes 30ul sample drops on slide, smear is carried out with another concave-concave piece,
Fluorecyte calculating instrument detects
Instrument title：Thermo Fisher, Countess II FL Automated Cell Counter.
Embodiment 3
1st, object peripheral blood blood sample 200ul to be measured is taken, adds antibody anti-CD14-FITC (BD companies, article No.
555397) 40ul, anti-CD16-APC (BD companies, article No. 561304) 10ul, room temperature lucifuge are incubated 15min；
2nd, add 200ul fixative room temperatures lucifuge and be incubated 5min；The composition of the fixative is：Percentage by volume is 1%
Formaldehyde, 0.35% methanol that percentage by volume is；
3rd, plus lysate 3mL is into sample, and the concussion that is vortexed mixes, and lucifuge is incubated 15min, splitting erythrocyte；The cracking
Liquid contains 10mM Tris-HCL, 10mM NaH2PO4And/or NaHPO4, 130mM NaCl, percentage by volume be 1% Triton
X-100、10mM Na4P2O7·10H2O, pH=7.5；
4th, 800g centrifuges 5min, removes supernatant；Supernatant slowly is outwelled, residual liquid is drawn with paper, the concussion that is vortexed is resuspended thin
Born of the same parents；
5th, 100ul closings and punching buffer solution are added, thrum mixes；
The Triton X-100 that the BSA and percentage by volume that closing and the punching buffer solution is 0.01g/mL are 1% are molten
Liquid；
6th, in above-mentioned solution system add fluorescence labeling anti-TKTL1 (antibody is purchased from rabbit polyclonal,
Sigma, article No. HPA000505, oneself adds fluorescence labeling) and the anti-DNASE1L1 antibody of fluorescence labeling (antibody is purchased from
Rabbit polyclonal, sigma, article No. HPA072635, oneself adds fluorescence labeling), thrum mixes, lucifuge incubation 15~
20min；The adding proportion of antibody is 1:1000；Pay attention to the anti-TKTL1 of fluorescence labeling and the anti-of fluorescence labeling
DNASE1L1 antibody should color and anti-CD14-FITC and anti-CD16-APC (BD companies, article No. 561304) it is glimmering
Light color is different.
7th, 2ml PBS (8g/L NaCl, 0.2g/L KCl, 1.4g/L Na are separately added into2HPO4、0.27g/L KH2PO4)
Wash, be vortexed and mix, 800g centrifugation 5min, remove supernatant, wash 2~3 times, cell is resuspended after washing；
8th, cell is resuspended with 500ul PBS, takes 30ul sample drops on slide, smear is carried out with another concave-concave piece,
Fluorecyte calculating instrument detects
Instrument title：Thermo Fisher, Countess II FL Automated Cell Counter.
Experimental example
1st, detected respectively with the method for embodiment 1~3 with three different samples (sample 1, sample 2, sample 3).
During statistics, first CD14 is found out from all monocytes+CD16+Cell and counting；Again from CD14+CD16+Cell in look for
Go out CD14+CD16+DNASE1L1+Or CD14+CD16+TKTL1+Cell and counting；Wherein, the way of control group is, by anti-
TKTL1 or anti-DNASE1L1 antibody replaces with the IgG antibody of same Species origin；Remaining step is consistent with experimental group.
Each embodiment testing result of table 1 (GCI values)
Wherein, detected value is respectively in form：
Control group：(CD14+CD16+IgG+÷CD14+CD16+)×1000；
DNASE1L1 groups：(CD14+CD16+DNASE1L1+÷CD14+CD16+)×1000；
TKTL1 groups：(CD14+CD16+TKTL1+÷CD14+CD16+)×1000；
End value is GCI (globle canceration index, global canceration index) value, and its algorithm is the inspection of each group
Measured value subtracts the detected value of control group.
Normal and disease line of demarcation value determines that the data of early stage are by substantial amounts of clinical data
TKTL1：It is normal less than 119, there is tumor risk more than 119 points；
DNASE1L1：0-100 is normal, and 100-130 suggests complete detection, and more than 130 have tumor risk；
Two groups of TKTL1 and DNASE1L1 of final result synthesis synthesis GCI values are considered.
2nd, same peripheral blood blood sample sample 4 is taken to be bisected into four groups, every group 3 parts, with embodiment 1~3 and comparative example
The method of record is detected, and every group in triplicate；The set-up mode of wherein comparative example is：Removed on the basis of embodiment 3
Add lysate splitting erythrocyte and centrifuge the step of going the removal of impurity, remaining operation is the same as embodiment 3 (ready-to-use whole blood is detected).Number
It is same as above according to computational methods.
The testing result of table 2 (GCI values)
As can be known from the above table, the method detection stability described in embodiment 1~3 is preferable and reproducible, and comparative example three
The floating of secondary detection is larger, and end value is far below embodiment, thus it is speculated that it is possible the reason for be due to not go the removal of impurity, antibody is non-
Specifically bind more, cause CD14+CD16+Value rises, and the antibody of tumor markers may be specific better, so
CD14+CD16+X+Change is little, and then causes GCI value entire lowerings, causes false negative.
Finally it should be noted that：Various embodiments above is merely illustrative of the technical solution of the present invention, rather than its limitations；To the greatest extent
The present invention is described in detail with reference to foregoing embodiments for pipe, but it will be understood by those within the art that：Its
The technical scheme described in foregoing embodiments can still be modified, either to which part or all technical characteristic
Carry out equivalent substitution；And these modifications or replacement, the essence of appropriate technical solution is departed from various embodiments of the present invention skill
The scope of art scheme.
Claims (10)
1. a kind of fluorecyte count detection kit for detecting tumor markers, it is characterised in that including marking cell
The CD14 antibody and CD16 antibody of film surface antigen, and to mark the antibody of endochylema internal antigens；
The endochylema internal antigens are TKTL1 and/or DNASE1L1；
The kit also includes microslide.
2. fluorecyte count detection kit according to claim 1, it is characterised in that the fluorecyte counts inspection
Test agent box is also including fluorescence corresponding to one or more antibody in the CD14 antibody, CD16 antibody and internal antigens antibody
Secondary antibody.
3. fluorecyte count detection kit according to claim 2, it is characterised in that the fluorescence secondary antibody is F
(ab') 2 antibody.
4. fluorecyte count detection kit according to claim 1, it is characterised in that the CD14 antibody, CD16
Antibody and internal antigens antibody are fluorescent labeled antibody, and fluorescence color is different.
5. fluorecyte count detection kit according to claim 1, it is characterised in that the fluorecyte counts inspection
Test agent box also includes the IgG antibody with the internal antigens antibody with Species origin.
6. the fluorecyte count detection kit according to any one of Claims 1 to 5, it is characterised in that the fluorescence
Cell count detection kit is also including any one in lysate, closing and punching buffer solution, fixer, rinsing liquid or more
Kind.
7. fluorecyte count detection kit according to claim 6, it is characterised in that the lysate be containing with
The working solution or its mother liquor of lower each composition of concentration：
8mM~12mM Tris-HCL, 8mM~12mM NaH2PO4And/or NaHPO4, 120mM~140mM NaCl, volume basis
Number is 0.7%~1.3% Triton X-100,8mM~12mM Na4P2O7·10H2O, pH=7.2~7.7.
8. fluorecyte count detection kit according to claim 6, it is characterised in that closing and the punching buffering
Liquid is working solution or its mother liquor containing each composition of following concentration：
0.008g/mL~0.012g/mL BSA and percentage by volume be 0.7%~1.3% TritonX-100 solution, solvent
For 1 × TBS or 1 × PBS solution.
9. fluorecyte count detection kit according to claim 6, it is characterised in that the fixer be containing with
The working solution or its mother liquor of lower each composition of concentration：
With volume percent, the mixed solution containing 0.8%~1.2% formaldehyde and 0.33%~0.38% methanol is used
Solvent is water.
10. fluorecyte count detection kit according to claim 6, it is characterised in that the rinsing liquid be containing
The working solution or its mother liquor of each composition of following concentration：
1 × PBS solution.
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| CN201610633992.0A CN107677822A (en) | 2016-08-02 | 2016-08-02 | Detect the fluorecyte count detection kit of tumor markers |
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| CN201610633992.0A CN107677822A (en) | 2016-08-02 | 2016-08-02 | Detect the fluorecyte count detection kit of tumor markers |
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|---|---|---|---|---|
| CN1726393A (en) * | 2002-12-18 | 2006-01-25 | 约翰内斯·科伊 | Compounds and methods for detection of carcinomas and their precursor lesions |
| CN101142486A (en) * | 2005-03-07 | 2008-03-12 | 约翰内斯·科伊 | Therapeutic and diagnostic uses of tktl1 and inhibitors and activators thereof |
| CN102914657A (en) * | 2012-10-10 | 2013-02-06 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Method for detecting protein in peripheral blood karyocyte cells |
| WO2013126872A1 (en) * | 2012-02-24 | 2013-08-29 | Wayne State University | Anti-cancer therapeutic strategy to overcome cancer resistance and to enable tailoring treatment to patients |
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2016
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|---|---|---|---|---|
| CN1726393A (en) * | 2002-12-18 | 2006-01-25 | 约翰内斯·科伊 | Compounds and methods for detection of carcinomas and their precursor lesions |
| CN101142486A (en) * | 2005-03-07 | 2008-03-12 | 约翰内斯·科伊 | Therapeutic and diagnostic uses of tktl1 and inhibitors and activators thereof |
| WO2013126872A1 (en) * | 2012-02-24 | 2013-08-29 | Wayne State University | Anti-cancer therapeutic strategy to overcome cancer resistance and to enable tailoring treatment to patients |
| CN102914657A (en) * | 2012-10-10 | 2013-02-06 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | Method for detecting protein in peripheral blood karyocyte cells |
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