CN104311664A - Preparation of fluorescent antibody of rabbit anti-human lung adenocarcinoma cells A549 - Google Patents
Preparation of fluorescent antibody of rabbit anti-human lung adenocarcinoma cells A549 Download PDFInfo
- Publication number
- CN104311664A CN104311664A CN201410593244.5A CN201410593244A CN104311664A CN 104311664 A CN104311664 A CN 104311664A CN 201410593244 A CN201410593244 A CN 201410593244A CN 104311664 A CN104311664 A CN 104311664A
- Authority
- CN
- China
- Prior art keywords
- antibody
- cell
- human lung
- lung adenocarcinoma
- rabbit
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention provides preparation of a fluorescent antibody of rabbit anti-human lung adenocarcinoma cells A549. Immunogen of human lung adenocarcinoma cells A549 is injected into a rabbit to prepare a polyclonal antibody of the rabbit anti-human lung adenocarcinoma cells A549. according to the invention, the adherent A549 cells are digested by pancreatin to prepare single-cell suspension, the cell density is adjusted, formaldehyde is added for inactivate, the A549 cell immunogen is subcutaneously injected in the rabbit (0.5 ml) at multipoint at the first, third, seventh and fourteenth days, and blood is extracted from the heart after immune injection of 30 days to separate the serum. IgG is roughly extracted by ammonium sulfate salt fractionation, IgG is purified by dextrangel, and the fluorescent antibody of rabbit anti-human lung adenocarcinoma cells A549 is prepared from a dialysis method. The antibody can be used for identifying multiple epitopes of the same antigen, improving the detection sensitivity under the same condition, can be used for verifying human lung adenocarcinoma cells and diagnosing immunopathogenesis of human lung adenocarcinoma, and has good social benefits and market prospects.
Description
Technical field
The present invention relates to technical field of pharmaceutical biotechnology, particularly relate to one and prepare the anti-human human umbilical vein endothelial cell fluorescence antibody of rabbit.
Background technology
Primary lung cancer is one of modal malignant tumour in the world, and it is the first that its case fatality rate occupies whole world cancer mortality reason; Its pathogenic factor comprises smoking, topsoil, some occupational factor, inherited genetic factors etc., and in the third time coroner's inquest of the whole nation, lung cancer accounts for 22.7% of the Nattonal Cancer cause of the death, is in the Nattonal Cancer cause of the death umber one.Lung cancer can be divided into nonsmall-cell lung cancer (NSCLC) and small cell lung cancer (SCLC) two class substantially, and wherein nonsmall-cell lung cancer accounts for the 80%-85% of lung cancer sum, and compared with small cell lung cancer, the growth division of its cancer cells is comparatively slow, and diffusion transfer is relatively late.
A549 cell belongs to NSCLC mono-class, derives from people's alveolar cell carcinoma, belongs to the adenocarcinoma cell in NSCLC classification, is set up by people such as Giard D J in 1973.This cell had both had the characteristic of lung cancer malignant cell, had again characteristic and the phenotype of type Ⅱ penumonocyte simultaneously.Therefore the dark welcome by lung cancer investigator, A549 cell is widely used in the generation of lung cancer, development, Treatment and diagnosis research, is also used to the research of the epithelial generation of normal people's alveolar type II, differentiation and structure and function thereof.According to Pubmed inquiry, up to the present, the English report using A549 cell to carry out studying is more than 6000 sections; The Chinese literature utilizing A549 cell to carry out studying that wherein CNKI includes reaches more than 4800 sections, and Sino-British document number rises in multiple level all year by year.
About the production of A549 monoclonal antibody, there is not been reported, and the A549 antibody of the present invention's application is this laboratory by by after A549 cellular immunization rabbit, extracts, polyclone IgG after purifying.
Fluorescein isothiocyanate (fluorescein isothiocyanate, FITC) be a kind of yellow, orange or brownish yellow powder or crystallization, stable in properties, soluble in water and alcohol, maximum absorption spectrum is 490-495nm, emission maximum spectrum is 520-530nm, presents yellow-green fluorescence, and molecular mass is 398.4.Under alkaline condition, the isothiocyanic acid base of FITC forms the amino key of sulphur carbon with the free amino group of immunoglobulin (Ig) through phosphinylidyne amination in aqueous, becomes mark fluorescent immune sphaeroprotein, i.e. fluorescence antibody, having good bonding force and stability, is the fluorescent marker that each laboratory often uses.
Immunofluorescence label technology (immunofluorescence technique) is by known antibody or the upper fluorescein of antigen molecule mark, when antigen corresponding thereto or antibody react, just with a certain amount of fluorescein on the mixture formed, under fluorescent microscope, just can see the antigen-antibody combining site sending fluorescence, detect antigen or antibody.
Summary of the invention
The object of the invention is to a kind of openly method preparing the anti-human human umbilical vein endothelial cell antibody of rabbit and immunofluorescence label, provide a kind of effective means directly perceived for lung cancer A549 detects.
Preparation rabbit anti-human human umbilical vein endothelial cell fluorescence antibody, its main preparation process comprises: immune animal in the cultivation of A549 cell, antigen preparation, body, extract the mark etc. of antibody, fluorescence antibody.
IgG prepared by the present invention is polyclonal antibody, can identify for multiple epi-positions of an antigen, improve the sensitivity detected, for lung cancer, there is no the good tumor associated antigen of generally acknowledged specificity clinically, the present invention can provide potential selection for the target inspection of lung cancer, for lung cancer laboratory diagnosis provides certain clinical value.
Technical scheme of the present invention and preparation process as follows:
1, A549 cell cultivation, go down to posterity: cell cultures, in 37 DEG C, puts 5%CO
2in incubator, under inverted microscope, observe A549 Growth of Cells
Form, until cell density close to 80% time, cell is carried out had digestive transfer culture.Nutrient solution old in bottle is outwelled, add 1ml pancreatin, outwelled rapidly after weak vibrations body, then added 1-2ml pancreatin, when observation of cell form change circle starts to come off under inverted microscope after 1-3min, add 4-5ml and stop digestion containing blood serum medium, after softly blowing and beating cell to cell suspension with suction pipe, go down to posterity with the ratio of 1:3, then the RPMI-1640 added containing 10%FBS is to 5ml, be statically placed in 37 DEG C, 5%CO
2cultivate in incubator.
2, the preparation of A549 antigen: by adherent A549 cell after trysinization, make single cell suspension, adjustment cell density is 10
6/ L, adds appropriate formalin-inactivated with the volume that the ratio of formaldehyde and cell is 1:10, places 4 DEG C of refrigerators for subsequent use.
3, immunity in animal body: immunologic process is the 1st, 3,7,14 day dorsal sc multi-point injection 0.5mlA549 cell, and immunization is Culling heart blood separation of serum after 30 days.
4, ammonium sulfate salting-out process slightly extracts IgG: immune serum 10 ml adds physiological saline 10 ml and mixes, and adds saturated ammonium sulphate 2Xml, (slowly drips while stirring, ammonium sulfate saturation ratio is made to reach 50%), 4 DEG C, leave standstill more than 3h, the centrifugal 20min of 3 000rpm, abandon supernatant, throw out normal saline dilution, to Xml, drips saturated coloured glaze acid ammonium 1/2 X ml while stirring, saturation ratio is made to be 33%, the centrifugal 20min of 4 DEG C of standing more than 3h, 3 000rpm, abandons supernatant.Repeat throw out normal saline dilution to Xml, drip saturated coloured glaze acid ammonium 1/2 X ml while stirring, make saturation ratio be the centrifugal 20min of 33%, 4 DEG C of standing more than 3h, 3 000rpm, abandon supernatant 2 times.Add a little physiological saline solution throw out for the last time and load dialysis tubing, put 4 DEG C and to dialyse desalination with PBS, change dialyzate for several times until: Nessler's reagent measures without NH
4 +, (have NH
4 +there will be brick-red or yellow), 1%BaCl
2measure without SO
4 2-(there is SO
4 2-there will be white precipitate), finally measure protein content (mg/ml), put 4 DEG C and save backup.
Measuring protein content method of calculation is:
IgG antibody protein content (mg/ml)=(1.45 × OD
280-0.74 × OD
260) × extension rate
IgG antibody titration double agar diffusion test Xiao Jia≤1:16, ELISA Shi Yan≤1:8000.
5, dextrane gel IgG purification: the pre-treatment of (1) gel: sephadex commodity mostly are dried particles, must be fully swelling before using.Method pours in the deionized water of 5 ~ 10 times lentamente by the xerogel for using, and heated and boiled method carries out gel swelling, and this method can not only accelerate swelling rate, and can remove the bacterium polluted in gel, gets rid of bubble simultaneously.(2) post is filled: the selection of chromatography column is generally determined according to the kind of sample separation and the quantity of sample.During protein purification, column volume should be 25 ~ 100 times of sample volume.The filling requirement of gel column, gel column requires that filler (gel) even density in post is consistent, does not have space and bubble.The gel column of usual new clothes is with after suitable buffered soln balance, by colored blue Pu Ju Tang – 2000, cytopigment, or the material such as oxyphorase to be mixed with mass concentration be that the solution of 2g/L crosses post, observe colour band whether evenly to move down, whether qualified to identify the technical quality of new clothes post, otherwise, must again load.(3) application of sample and wash-out: 1) application of sample amount: application of sample amount is relevant with chromatography column size with measuring method.Adopt 280nm wavelength to measure absorbancy, the post of a 2cm × 60cm, application of sample amount needs about 5mg.Less or injection volume less (sample concentration is high), resolving power is higher for application of sample amount.The add-on of usual sample liquid should rest in 5% ~ 10% of gel bed cumulative volume.Sample volume is excessive, and separating effect is bad.2) loading methods: gel bed, after balance, sucks supernatant liquid, when ready to balance liquid drops to bed surface, closes spout, adds sample liquid, open spout, sample liquid is slowly infiltrated in gel bed with dropper.When sample liquid level is proper and gel bed surface maintains an equal level, carefully adds several mL elutriant and rinse tube wall.Then continue to use a large amount of elution.3) wash-out: after adding sample, is connected chromatography bed with elutriant storage bottle, detector, fraction collector and registering instrument, and according to the character of separated material, pre-estimate what a suitable flow velocity, fraction collection effluent liquid quantitatively, every component one is to several mL.Each component can carry out qualitative or quantitative analysis by appropriate means.Gel filtration chromatography is general all using single buffered soln or salts solution as elutriant, sometimes even can use distilled water.The device of flow rate control is used for it is preferred that constant flow pump during wash-out.If without this device, the way of available red-tape operati pressure is carried out.Sample volume is too much unsuitable, is preferably 1% ~ 5% of bed volume, does not exceed 10% at most.Sample concentration is also unsuitable excessive, and the excessive viscosity of concentration is large, and inferior separating effect, is generally no more than 4%.Elutriant should be consistent with expansion, otherwise change solvent, and gel volume can change, and affects separating effect.Elutriant will have certain ionic strength and pH value.Separating serum proteins commonly uses PBS liquid (0.14Mol/L NaCl) and the 0.1Mol/L pH8.0Tris-HCl buffer salt solution (0.14Mol/L NaCl) of 0.02 ~ 0.1Mol/L pH 6.9 ~ 8.0.6, A549 antibody fluorescence mark: according to for labelled protein total amount, in FITC: albumen is the ratio of 1:10, accurately take required FITC powder with analytical balance.FITC powder is inserted in beaker, adds a certain amount of carbonate buffer solution, make final protein concentration be 20mg/ml, mixing.Be diluted to 10mg/ml for traget antibody carbonate buffer solution, get the dialysis tubing cleaned up, one end ties up, and adds the antibody diluted in right amount, tie up the other end from the other end.Being inserted by dialysis tubing is equipped with in the beaker of FITC, at 4 DEG C, and magnetic stirrer 24 hours.
7, fluorescence antibody dialysis: antibody labeling is after 24 hours, take out dialysis tubing and put into phosphoric acid buffer and dialyse, every day changes liquid 2-4 time, removes non-specific pigment, continuous 6-7 days, until dialyzate under uviolizing till complete unstressed configuration.
8, the preservation of fluorescence antibody: with 0-4 DEG C or-20 DEG C of cryopreservation, prevents antibody activity from reducing or sex change, can add the NaN of concentration for (1:5000)-(1:10000)
3, packing in a small amount, avoids multigelation.
9, feature of the present invention is: this antibody is polyclonal antibody, can identify multiple epi-positions of same antigen, and under the same conditions, can improve the sensitivity of detection.
Embodiment
The specific embodiment of the present invention, is now presented below its technical scheme and step again:
1, A549 cell cultivation, go down to posterity: cell cultures, in 37 DEG C, puts 5%CO
2in incubator, under inverted microscope, observe A549 Growth of Cells
Form, until cell density close to 80% time, cell is carried out had digestive transfer culture.Nutrient solution old in bottle is outwelled, add 1ml pancreatin, outwelled rapidly after weak vibrations body, then added 1-2ml pancreatin, when observation of cell form change circle starts to come off under inverted microscope after 1-3min, add 4-5ml and stop digestion containing blood serum medium, after softly blowing and beating cell to cell suspension with suction pipe, go down to posterity with the ratio of 1:3, then the RPMI-1640 added containing 10%FBS is to 5ml, be statically placed in 37 DEG C, 5%CO
2cultivate in incubator.
2, the preparation of A549 antigen: by adherent A549 cell after trysinization, make single cell suspension, adjustment cell density is 10
6/ L, adds appropriate formalin-inactivated with the volume that the ratio of formaldehyde and cell is 1:10, places 4 DEG C of refrigerators for subsequent use.
3, immunity in animal body: immunologic process is the 1st, 3,7,14 day dorsal sc multi-point injection 0.5mlA549 cell, and immunization is Culling heart blood separation of serum after 30 days.
4, ammonium sulfate salting-out process slightly extracts IgG: immune serum 10 ml adds physiological saline 10 ml and mixes, and adds saturated ammonium sulphate 2Xml, (slowly drips while stirring, ammonium sulfate saturation ratio is made to reach 50%), 4 DEG C, leave standstill more than 3h, the centrifugal 20min of 3 000rpm, abandon supernatant, throw out normal saline dilution, to Xml, drips saturated coloured glaze acid ammonium 1/2 X ml while stirring, saturation ratio is made to be 33%, the centrifugal 20min of 4 DEG C of standing more than 3h, 3 000rpm, abandons supernatant.Repeat throw out normal saline dilution to Xml, drip saturated coloured glaze acid ammonium 1/2 X ml while stirring, make saturation ratio be the centrifugal 20min of 33%, 4 DEG C of standing more than 3h, 3 000rpm, abandon supernatant 2 times.Add a little physiological saline solution throw out for the last time and load dialysis tubing, put 4 DEG C and to dialyse desalination with PBS, change dialyzate for several times until: Nessler's reagent measures without NH
4 +, (have NH
4 +there will be brick-red or yellow), 1%BaCl
2measure without SO
4 2-(there is SO
4 2-there will be white precipitate), finally measure protein content (mg/ml), put 4 DEG C and save backup.
Measuring protein content method of calculation is:
IgG antibody protein content (mg/ml)=(1.45 × OD
280-0.74 × OD
260) × extension rate
IgG antibody titration double agar diffusion test Xiao Jia≤1:16, ELISA Shi Yan≤1:8000.
5, dextrane gel IgG purification: the 1) pre-treatment of gel: sephadex commodity mostly are dried particles, must be fully swelling before using.Method pours in the deionized water of 5 ~ 10 times lentamente by the xerogel for using, and heated and boiled method carries out gel swelling, and this method can not only accelerate swelling rate, and can remove the bacterium polluted in gel, gets rid of bubble simultaneously.2) post is filled: the selection of chromatography column is generally determined according to the kind of sample separation and the quantity of sample.During protein purification, column volume should be 25 ~ 100 times of sample volume.The filling requirement of gel column, gel column requires that filler (gel) even density in post is consistent, does not have space and bubble.The gel column of usual new clothes is with after suitable buffered soln balance, by colored blue Pu Ju Tang – 2000, cytopigment, or the material such as oxyphorase to be mixed with mass concentration be that the solution of 2g/L crosses post, observe colour band whether evenly to move down, whether qualified to identify the technical quality of new clothes post, otherwise, must again load.3) application of sample and wash-out: 1. application of sample amount: application of sample amount is relevant with chromatography column size with measuring method.Adopt 280nm wavelength to measure absorbancy, the post of a 2cm × 60cm, application of sample amount needs about 5mg.Less or injection volume less (sample concentration is high), resolving power is higher for application of sample amount.The add-on of usual sample liquid should rest in 5% ~ 10% of gel bed cumulative volume.Sample volume is excessive, and separating effect is bad.2. loading methods: gel bed, after balance, sucks supernatant liquid, when ready to balance liquid drops to bed surface, closes spout, adds sample liquid, open spout, sample liquid is slowly infiltrated in gel bed with dropper.When sample liquid level is proper and gel bed surface maintains an equal level, carefully adds several mL elutriant and rinse tube wall.Then continue to use a large amount of elution.3. wash-out: after adding sample, is connected chromatography bed with elutriant storage bottle, detector, fraction collector and registering instrument, and according to the character of separated material, pre-estimate what a suitable flow velocity, fraction collection effluent liquid quantitatively, every component one is to several mL.Each component can carry out qualitative or quantitative analysis by appropriate means.Gel filtration chromatography is general all using single buffered soln or salts solution as elutriant, sometimes even can use distilled water.The device of flow rate control is used for it is preferred that constant flow pump during wash-out.If without this device, the way of available red-tape operati pressure is carried out.Sample volume is too much unsuitable, is preferably 1% ~ 5% of bed volume, does not exceed 10% at most.Sample concentration is also unsuitable excessive, and the excessive viscosity of concentration is large, and inferior separating effect, is generally no more than 4%.Elutriant should be consistent with expansion, otherwise change solvent, and gel volume can change, and affects separating effect.Elutriant will have certain ionic strength and pH value.Separating serum proteins commonly uses PBS liquid (0.14Mol/L NaCl) and the 0.1Mol/L pH8.0Tris-HCl buffer salt solution (0.14Mol/L NaCl) of 0.02 ~ 0.1Mol/L pH 6.9 ~ 8.0.6, A549 antibody fluorescence mark: according to for labelled protein total amount, in FITC: albumen is the ratio of 1:10, accurately take required FITC powder with analytical balance.FITC powder is inserted in beaker, adds a certain amount of carbonate buffer solution, make final protein concentration be 20mg/ml, mixing.Be diluted to 10mg/ml for traget antibody carbonate buffer solution, get the dialysis tubing cleaned up, one end ties up, and adds the antibody diluted in right amount, tie up the other end from the other end.Being inserted by dialysis tubing is equipped with in the beaker of FITC, at 4 DEG C, and magnetic stirrer 24 hours.
7, fluorescence antibody dialysis: antibody labeling is after 24 hours, take out dialysis tubing and put into phosphoric acid buffer and dialyse, every day changes liquid 2-4 time, removes non-specific pigment, continuous 6-7 days, until dialyzate under uviolizing till complete unstressed configuration.
8, the preservation of fluorescence antibody: with 0-4 DEG C or-20 DEG C of cryopreservation, prevents antibody activity from reducing or sex change, can add the NaN of concentration for (1:5000)-(1:10000)
3, packing in a small amount, avoids multigelation.
9, this antibody is polyclonal antibody, can identify multiple epi-positions of same antigen, and under the same conditions, can improve the sensitivity of detection.
Claims (2)
1. prepare the anti-human human umbilical vein endothelial cell fluorescence antibody of rabbit, it is characterized in that: it comprises the steps:
1) A549 cell cultivation, go down to posterity: cell cultures, in 37 DEG C, puts 5%CO
2in incubator, cell density close to 80% time, cell is carried out had digestive transfer culture;
2) preparation of A549 antigen: by adherent A549 cell after trysinization, make single cell suspension, adjustment cell density is 10
6/ L, adds appropriate formalin-inactivated with the volume that the ratio of formaldehyde and cell is 1:10, places 4 DEG C of refrigerators for subsequent use;
3) immunity in animal body: immunologic process be the 1st, 3,7,14 day in rabbit dorsal sc multi-point injection 0.5mlA549 cell, immunization is Culling heart blood separation of serum after 30 days;
4) ammonium sulfate salting-out process slightly extracts IgG: immune serum 10 ml adds physiological saline 10 ml and mixes, add saturated ammonium sulphate 2Xml, (slowly drip while stirring, make ammonium sulfate saturation ratio reach 50%), 4 DEG C, leave standstill the centrifugal 20min of more than 3h, 3 000rpm, abandon supernatant, throw out normal saline dilution is to Xml, drip saturated coloured glaze acid ammonium 1/2 X ml while stirring, make saturation ratio be 33%, repeat 2 times;
Add a little physiological saline solution throw out for the last time and load dialysis tubing, put 4 DEG C and to dialyse desalination with PBS, change dialyzate for several times until: Nessler's reagent measures without NH
4 +, have NH
4 +there will be brick-red or yellow, 1%BaCl
2measure without SO
4 2-, have SO
4 2-there will be white precipitate, finally measure protein content mg/ml, put 4 DEG C and save backup;
IgG antibody titration double agar diffusion test Xiao Jia≤1:16, ELISA Shi Yan≤1:8000;
5) dextrane gel IgG purification: pour in the deionized water of 5 ~ 10 times lentamente by the xerogel for using, heated and boiled method carries out gel swelling; Dress post: should be 25 ~ 100 times of sample volume by column volume; Application of sample and wash-out adopt 280nm wavelength to measure absorbancy, the post of a 2cm × 60cm, and application of sample amount needs about 5mg; After adding sample, chromatography bed is connected with elutriant storage bottle, detector, fraction collector and registering instrument, be separated the PBS liquid with 0.02 ~ 0.1Mol/L pH 6.9 ~ 8.0, i.e. 0.14Mol/L NaCl and 0.1Mol/L pH8.0Tris-HCl buffer salt solution, i.e. 0.14Mol/L NaCl;
6) A549 antibody fluorescence mark: in FITC: albumen is the ratio of 1:10, accurately take required FITC powder with analytical balance; FITC powder is inserted in beaker, adds a certain amount of carbonate buffer solution, make final protein concentration be 20mg/ml, mixing; Be diluted to 10mg/ml for traget antibody carbonate buffer solution, get the dialysis tubing cleaned up, one end ties up, and adds the antibody diluted in right amount, tie up the other end from the other end; Being inserted by dialysis tubing is equipped with in the beaker of FITC, at 4 DEG C, and magnetic stirrer 24 hours;
7) fluorescence antibody dialysis: antibody labeling is after 24 hours, take out dialysis tubing and put into phosphoric acid buffer and dialyse, every day changes liquid 2-4 time, removes non-specific pigment, continuous 6-7 days, until dialyzate under uviolizing till complete unstressed configuration;
8) preservation of fluorescence antibody: with 0-4 DEG C or-20 DEG C of cryopreservation, prevents antibody activity from reducing or sex change, can add the NaN that concentration is 1:5000-1:10000
3, packing in a small amount, avoids multigelation.
2. the anti-human human umbilical vein endothelial cell fluorescence antibody of preparation rabbit as claimed in claim 1, is characterized in that: step 2) cell density of preparing of A549 antigen is 10
6/ L, adds formalin-inactivated with the volume that the ratio of formaldehyde and cell is 1:10; 3) immunity in animal body: immunologic process be the 1st, 3,7,14 day in rabbit dorsal sc multi-point injection 0.5mlA549 cell, immunization is Culling heart blood separation of serum after 30 days; 4) ammonium sulfate salting-out process slightly extracts IgG titration double agar diffusion test Xiao Jia≤1:16, and ELISA Shi Yan≤1:8000 is qualified; 6) A549 antibody fluorescence mark: in FITC: albumen is the ratio of 1:10, make final protein concentration be 20mg/ml, mixing; Be diluted to 10mg/ml for traget antibody carbonate buffer solution, get the dialysis tubing cleaned up, one end ties up, from the other end, add the antibody diluted in right amount, tie up the other end, dialysis tubing is inserted and is equipped with in the beaker of FITC, at 4 DEG C, magnetic stirrer 24 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410593244.5A CN104311664A (en) | 2014-10-30 | 2014-10-30 | Preparation of fluorescent antibody of rabbit anti-human lung adenocarcinoma cells A549 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410593244.5A CN104311664A (en) | 2014-10-30 | 2014-10-30 | Preparation of fluorescent antibody of rabbit anti-human lung adenocarcinoma cells A549 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104311664A true CN104311664A (en) | 2015-01-28 |
Family
ID=52366996
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410593244.5A Pending CN104311664A (en) | 2014-10-30 | 2014-10-30 | Preparation of fluorescent antibody of rabbit anti-human lung adenocarcinoma cells A549 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104311664A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101339192A (en) * | 2008-08-28 | 2009-01-07 | 河南省农业科学院 | Test paper for one-step detection for pig virus diarrhoea disease pathogen |
| CN101484464A (en) * | 2006-02-17 | 2009-07-15 | 诺华有限公司 | Purification of bacterial antigens |
-
2014
- 2014-10-30 CN CN201410593244.5A patent/CN104311664A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101484464A (en) * | 2006-02-17 | 2009-07-15 | 诺华有限公司 | Purification of bacterial antigens |
| CN101339192A (en) * | 2008-08-28 | 2009-01-07 | 河南省农业科学院 | Test paper for one-step detection for pig virus diarrhoea disease pathogen |
Non-Patent Citations (2)
| Title |
|---|
| 母波等: "兔抗人多发性骨髓瘤细胞多克隆抗体的制备与鉴定", 《江西科学》 * |
| 阚兵: "抗人肺癌单克隆抗体的制备及其生物特性的研究", 《四川大学硕士学位论文》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111303254A (en) | Novel coronavirus (SARS-CoV-2) antigen detection kit | |
| CN103787946B (en) | NSC 70328 artificial antigen and antibody and its preparation method and application | |
| CN103820394B (en) | Monoclonal antibodies against morphine, produce the cell strain of this antibody, morphine detection kit and preparation method thereof | |
| CN104316703B (en) | A kind of Mycoplasma bovis test strip and its preparation method | |
| CN104792997A (en) | Human procalcitonin immunodetection kit, and preparation method and application thereof | |
| CN103575889A (en) | Test strip and method for detecting vancomycin | |
| CN105950563A (en) | Hybridoma cell strain 7E3, monoclonal antibody secreted by hybridoma cell strain 7E3 and resistant to FMD type A virus, and application | |
| CN103333864B (en) | Monoclonal antibody of toxoplasma gondii resistant MIC3 protein and application monoclonal antibody | |
| CN105759043A (en) | Test paper for simultaneous detection of carbendazim and thiophanate-methyl residues in crops, and application and preparation method thereof | |
| CN105542014A (en) | TP recombinant antigen and preparing method and application thereof | |
| CN103698536B (en) | Oncoprotein P185 detection kit | |
| CN101144819B (en) | Colloidal gold test paper for detecting sulphaquinoxaline drug residue | |
| CN105445462A (en) | Enzyme linked immunosorbent assay kit for specific quantitative detection of foot and mouth disease O type Guangxi strain antigen and application thereof | |
| CN103739675B (en) | Strawberry veinbanding virus antibody and antigenic peptide, immunogen and application | |
| CN103389379A (en) | Test strip and method for detecting tylosin and tilmicosin | |
| CN107014992B (en) | A kind of foundation of quantitative detection human blood type antibody concentration method | |
| CN106399294B (en) | A kind of preparation of the monoclonal antibody 7H8 of anti-human Procalcitonin protein N terminal epitope | |
| CN105732810A (en) | Procalcitonin monoclonal antibody and application thereof | |
| CN106957362A (en) | A kind of preparation method and applications of Pichia pastoris host protein antibody | |
| CN107746430B (en) | Preparation and application of GP 73C-terminal antigen | |
| CN102183641A (en) | Ractopamine immunochromatographic assay test paper strip | |
| CN105481957A (en) | Platanus pollen allergen Pla a 3 and monoclonal antibody thereof | |
| CN104311664A (en) | Preparation of fluorescent antibody of rabbit anti-human lung adenocarcinoma cells A549 | |
| CN102297964A (en) | Immunity chromatography detection test strip of melamine | |
| CN101196523A (en) | Chicken infectious Fabricius bursa immune body immune colloidal gold fast detecting and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150128 |
|
| WD01 | Invention patent application deemed withdrawn after publication |