CN104422659B - The application of salicylide azine and its derivative in protein fluorescence detection - Google Patents
The application of salicylide azine and its derivative in protein fluorescence detection Download PDFInfo
- Publication number
- CN104422659B CN104422659B CN201310268922.6A CN201310268922A CN104422659B CN 104422659 B CN104422659 B CN 104422659B CN 201310268922 A CN201310268922 A CN 201310268922A CN 104422659 B CN104422659 B CN 104422659B
- Authority
- CN
- China
- Prior art keywords
- application
- azine
- salicylide
- protein
- concentration
- 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.)
- Active
Links
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention relates to protein fluorescence detection technology, the specifically application of salicylide azine (Salicylaldehyde azine) and its derivative in protein fluorescence detection.Present invention also offers the method that application salicylide azine carries out protein fluorescence detection, including step：Gel after protein example electrophoresis is fixed in fixer, deionized water rinsing；Dyeing；Pickling；Alkali lye decolourizes；Pickling；Detection.The present invention have sensitivity it is high, it is simple to operate it is rapid, favorable reproducibility, dyeing background are good, linear relationship is good, good reversibility, compatibility is good, use safety, low cost and other advantages.
Description
Technical field
The present invention relates to protein fluorescence detection technology, a kind of new fluorescent dye of protein is related in particular to.
Background technology
In life science field, the base substance protein to life and DNA research are be unable to do without, is also just be unable to do without
To the detection technique of protein.Especially recent years, with developing rapidly for genomics and proteomics, make us
There is higher requirement to the separation detection technique of protein.
The detection method of protein has a lot, including nephelometry, Kjeldahl's method, absorption photometry, fluorimetry, with
And resonance rayleigh light scattering method, high performance liquid chromatography, the capillary electrophoresis analysis method that new development is got up.Fluorimetry is mesh
Preceding most study, most widely used, operation detection method relatively simple, in higher sensitivity.
Salicylic acid azine and its derivative are applied to protein fluorescence detection by the present invention first, compared to the inspection of other fluorescence
Survey technology, with many merits, will be with a wide range of applications in proteomics research.
The content of the invention
It is an object of the invention to set up a kind of application of salicylide azine and its derivative in protein fluorescence detection.
Salicylide azine related compound of the present invention refers to sodium salt, sylvite, the salt using salicylide azine as parent nucleus
Hydrochlorate, sulfate, disulfate, nitrate, tannate, hydriodate etc..
Salicylide azine parent nucleus is：
Here is an available Gel Detection Method of the invention, and it includes step：
1) the protein example gel after SDS-PAGE electrophoresis is placed in fixer and fixes 10~60min, abandon fixation
Liquid, then with 1~20min of deionized water rinsing；It is preferred that set time be 20min, the deionized water rinsing time be 5min
(totally 2 times), fixer can be the aqueous solution containing 40% ethanol and 10% acetic acid；
2) add fluorescent staining liquid and dye 1~30min, 1~10min wash with pickle, wherein the dyeing liquor for containing
By 25~50 μM salicylide azine of molar concentration or derivatives thereof and by the aqueous solution of molar concentration 5~15mM sodium hydroxides,
The acetic acid that it is by volume 0.01~1% that pickle, which is,；The concentration of preferred salicylide azine or derivatives thereof is in dyeing liquor
25 μM, preferred naoh concentration is 10mM, and preferred dyeing time is that preferred acetic acid concentration in 5min, pickle is
0.1%, pickling time preferably is 5min；
3) add alkali wash water and wash 5~30min, wherein the alkali lye is the borax containing volume ratio 0.1~1% by weight,
0.1~2% sodium carbonate；It is preferred that borate concentration be 0.2%, preferred concentration of sodium carbonate is 0.5%, during preferred washing
Between be 20min；
4) add terminate liquid and wash 5~30min, wherein the terminate liquid is the second containing volume ratio 0.01~1% by weight
Acid；It is preferred that acetic acid concentration be 0.1%, preferred termination time is 5min；
5) detect, Detection wavelength is 250~370nm, preferably 312nm.
The inventive method has the following advantages that：
1) high sensitivity：More much higher than the sensitivity of common dye times of fluorescent dye；
2) it is simple to operate rapid：Operating procedure is few, can be completed within half an hour；
3) favorable reproducibility：Influenceed smaller by external conditions such as temperature, shaking table hunting frequencies；
4) dyeing background is good；
5) linear relationship is good：Semiquantitative determination can be carried out to protein interior in a big way；
6) good reversibility：Easily decolourize；
7) compatibility is good：Can be with the mass spectrometer highly compatible such as MALDI-TOF；
8) using safety：Using toxicity very low natural fluorescent dye, the security of experimental implementation, environmental pollution are improved
It is small；
9) cost is low.
Brief description of the drawings
The chemical constitution of Fig. 1 salicylide azines；
Fig. 2 salicylide azine fluorescence colours and the comparison of other conventional decoration method sensitivity.Wherein (A) is salicylide
Azine decoration method, (B) is EY negative stainings, and (C) dyes for SYPRO-Ruby, and (D) is glutaraldehyde argentation.Protein is commodity
Change standard items, from top to bottom respectively transferrin (transferrins), bovine serum albumin(BSA) (bovine serum
Albumin), immunoglobulin (IgG), ovalbumin (OVA), α1- acidoglycoprotein (α1- acid glycoprotein), α-
Casein (α-casein), beta-casein (β-casein), κ-casein (κ-casein), avidin (avidin).Protein
Volume containing the sample (per band) is as follows：Band 1,100ng；Band 2,50ng；Band 3,25ng；Band 4,12.5ng；Band 5,6.4ng；Band 6,
3.2ng；Band 7,1.6ng；Band 8,0.8ng；Band 9,0.4ng；Band 10,0.2ng；
Fig. 3 salicylide azine fluorescence colours and the comparison of other conventional decoration method sensitivity.Wherein (A) is salicylide
Azine decoration method, (B) is EY negative stainings, and (C) dyes for SYPRO-Ruby, and (D) is glutaraldehyde argentation, and protein is large intestine
Bacillus holoprotein, (1-10) times times dilution from left to right of protein volume containing the sample.
Embodiment：
The present invention is expanded on further with reference to specific embodiment.It should be appreciated that these embodiments are merely to illustrate
The present invention, and can not limit the scope of the invention.
The salicylide azine of embodiment 1 is dyed
Fig. 1 is the chemical structural formula of salicylide azine.Salicylide azine protein fluorescence Coloration experiment uses following step
Carry out：
1) protein example (Sigma-Aldrich Co, USA) gel after SDS-PAGE electrophoresis is placed in fixer
Middle 20min, twice, each 5min, fixer is the ethanol of volumn concentration 40%, the water of 10% acetic acid to deionized water rinsing
Solution；
2) 5min is dyed in dyeing liquor, 5min is washed with 0.1% acetic acid, dyeing liquor be containing 25 μM of salicylide azines and
The aqueous solution of 10mM sodium hydroxides；
3) 20min is washed in alkali wash water, alkali wash water is containing 0.2% borax, the aqueous solution of 0.5% sodium carbonate.
4) 5min is terminated in acid solution, terminate liquid is containing 0.1% acetic acid aqueous solution.
5) gel being colored is placed directly on transilluminator, is transmitted at 312nm, recording image.
Use sodium salt, sylvite, hydrochloride, sulfate, disulfate, the nitric acid of salicylide azine respectively according to the method described above
Salt, tannate, hydriodate carry out protein staining, as a result show be obtained being similar to salicylide a word used for translation with these derivatives
The testing result of piperazine.SDS-PAGE references《Molecular Cloning:A Laboratory guide》Third edition relevant portion is carried out.
The salicylide azine of experimental example 1 and other dyeing Contrast on effect
Dyed using different dyes, (A) is salicylide azine decoration method, (B) is EY negative stainings, and (C) is SYPRO-
Ruby is dyed, and (D) is glutaraldehyde argentation.Protein volume containing the sample (per band) is as follows：Band 1,100ng；Band 2,50ng；Band 3,
25ng；Band 4,12.5ng；Band 5,6.4ng；Band 6,3.2ng；Band 7,1.6ng；The ng of band 8,0.8；Band 9,0.4ng；Band 10,
0.2ng.As a result as shown in Fig. 2 salicylide azine decoration method remolding sensitivity SYPRO-Ruby is dyed, glutaraldehyde argentation is high, with
EY negative stainings are close；But in small-molecular-weight region, salicylide azine decoration method remolding sensitivity EY negative stainings are high.Wherein bigcatkin willow
The dyeing of aldehyde azine is carried out using the method for embodiment 1, and EY negative staining, SYPRO-Ruby dyeing and glutaraldehyde argentation are pressed respectively
The mode of stating is carried out：
EY negative staining[1]：
1) gel after electrophoresis is placed in 20min in fixer (50% ethanol, 10% acetic acid)；
2) deionized water washing 5min, totally 2 times；
3) gel is placed in 50% equilibrium methanol liquid and balances 10min；
4) the dyeing 15min in dyeing liquor (0.4%Eosin Y, 50% methanol, 1% acetic acid), then washed with deionized water
3min；
5) glue being colored is placed on recording image or direct dry glue on light-passing board.
SYPRO Ruby[2]Decoration method：
1) gel after electrophoresis is placed on into dyeing in SYPRO Ruby dyeing working solutions to be no less than 3 hours, then uses 10% first
The acetic acid solution of alcohol/7% rinses 30min；
2) gel being colored is placed directly on transilluminator, is transmitted at 312nm, recording image.
Glutaraldehyde argentation[3]：
1) gel after electrophoresis is placed in 30min in fixer (40% ethanol, 10% acetic acid)；
2) gel is placed in 6.8% sodium acetate, 0.125% glutaraldehyde, the reaction solution of 0.2% sodium thiosulfate and reacted
30min；
3) 5min, totally 3 times are washed with deionized water；
4) gel is placed in dyeing 20min in dyeing liquor (0.015% formaldehyde, 0.25% silver nitrate)；
5) 1min, totally 2 times are washed with deionized water；
6) gel is placed in developer solution (3% sodium carbonate, 0.007% formaldehyde) and developed；
7) gel is placed in reaction 10min in terminate liquid (1.5%EDTA).
Bibliography：
1.Wei-Tao Cong, Sun-Young Hwang, High-throughput negative detection
ofSDS-PAGE separated proteins and its application for
Proteomics.Electrophoresis 2010,31,411-420.
2.Berggren, K., Chernokalskaya, E., Steinberg, T.H., Kemper, C., Lopez, M.F.,
Diwu, Z., Haugland, R.P., Patton, W.F., Electrophoresis 2000,21,2509-2521.
3.Heukeshoven, J., Dernick, R., Electrophoresis 1985,6,103-112.
Claims (9)
1. the derivative of salicylide azine or salicylide azine is as the fluorescent dye of protein after SDS-PAGE electrophoresis
Application in the fluoroscopic examination of protein in protein example gel, wherein, the salicylide azine derivatives are salicylides
Sodium salt, sylvite, hydrochloride, sulfate, disulfate, nitrate, tannate or the hydriodate of azine.
2. the application described in claim 1, wherein the step of fluoroscopic examination is：
1) the protein example gel after SDS-PAGE electrophoresis is placed in fixer and fixes 10~60min, abandon fixer, so
1~20min of deionized water rinsing, altogether 2 times are used afterwards；
2) add fluorescent staining liquid and dye 1~30min, wherein the dyeing liquor is containing the salicylide by 25~50 μM of molar concentration
The derivative and the aqueous solution by molar concentration 5~15mM sodium hydroxides of azine or salicylide azine；
3) add pickle and wash 5~30min, wherein the pickle is the acetic acid containing volume ratio 0.01~1% by weight；
4) add alkali wash water and wash 20~40min, wherein the alkali wash water is the borax containing volume ratio 0.1~1% by weight,
0.1~2% sodium carbonate；
5) add terminate liquid and wash 5~30min, wherein the terminate liquid is the acetic acid containing volume ratio 0.01~1% by weight；
6) detect.
3. application as claimed in claim 2, it is characterised in that step 1) in the proteopexy time be 20min, then spend from
Sub- water rinses 5min, totally 2 times.
4. application as claimed in claim 2, it is characterised in that step 2) in time of dyeing be 5min.
5. application as claimed in claim 2, it is characterised in that step 2) in dyeing liquor salicylide azine concentration by mole dense
Spend for 25 μM, naoh concentration is 10mM in dyeing liquor.
6. method as claimed in claim 2, it is characterised in that step 3) the bulking value specific concentration of acetic acid is in pickle
0.1%.
7. application as claimed in claim 2, it is characterised in that step 4) the bulking value specific concentration of borax is in alkali wash water
0.2%, the bulking value specific concentration of sodium carbonate is 0.5%.
8. application as claimed in claim 2, it is characterised in that step 4) the alkali cleaning time is 20min.
9. application as claimed in claim 2, it is characterised in that step 5) the bulking value specific concentration of acetic acid is in terminate liquid
0.1%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310268922.6A CN104422659B (en) | 2013-08-29 | 2013-08-29 | The application of salicylide azine and its derivative in protein fluorescence detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310268922.6A CN104422659B (en) | 2013-08-29 | 2013-08-29 | The application of salicylide azine and its derivative in protein fluorescence detection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104422659A CN104422659A (en) | 2015-03-18 |
| CN104422659B true CN104422659B (en) | 2017-09-19 |
Family
ID=52972282
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310268922.6A Active CN104422659B (en) | 2013-08-29 | 2013-08-29 | The application of salicylide azine and its derivative in protein fluorescence detection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104422659B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105445244B (en) * | 2015-11-13 | 2018-08-28 | 南京林业大学 | A method of measuring potassium tert-butoxide content |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1858574A (en) * | 2006-04-04 | 2006-11-08 | 中国药科大学 | Method for synchronously dyeing protein and polyethylene glycol on electrophoretic gel |
| CN101113983A (en) * | 2007-08-30 | 2008-01-30 | 温州医学院 | Application of Palmatine and its derivatives in protein fluorescent detecting |
| CN201803885U (en) * | 2010-09-13 | 2011-04-20 | 深圳市卫武光明生物制品有限公司 | Gel dyeing and decolorizing device |
| CN102053030A (en) * | 2009-11-03 | 2011-05-11 | 广东暨大基因药物工程研究中心有限公司 | Mass spectrum-compatible protein argentation |
| CN102061632A (en) * | 2010-11-19 | 2011-05-18 | 南京工业大学 | Dyeing process of natural organic dye extracted from safflower |
| CN102200526A (en) * | 2011-03-15 | 2011-09-28 | 六安初创人乳库有限公司 | Qualitative detection method capable of distinguishing cow milk doped in human milk |
| CN102590099A (en) * | 2011-01-18 | 2012-07-18 | 温州医学院 | Application of 8-phenylamino-1-naphthalene sulfonic acid and derivative thereof to protein fluorescence detection |
| CN102607920A (en) * | 2012-03-15 | 2012-07-25 | 中国热带农业科学院热带生物技术研究所 | SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) coomassie brilliant blue R250 fast staining solution, staining method and application |
| CN102911189A (en) * | 2012-10-16 | 2013-02-06 | 桂林理工大学 | Salicylaldehyde azine schiff base transition metal compound and application thereof |
| CN103149262A (en) * | 2013-01-11 | 2013-06-12 | 安徽中医学院第一附属医院 | Xinfeng capsule water-soluble protein molecular weight detecting method |
| CN103233060A (en) * | 2013-04-26 | 2013-08-07 | 天津科技大学 | Method for analyzing protein expression during fungal fermentation process |
| CN103267666A (en) * | 2013-02-21 | 2013-08-28 | 山东理工大学 | Protein and gamma-polyglutamic acid simultaneous coloring counterstaining method |
-
2013
- 2013-08-29 CN CN201310268922.6A patent/CN104422659B/en active Active
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1858574A (en) * | 2006-04-04 | 2006-11-08 | 中国药科大学 | Method for synchronously dyeing protein and polyethylene glycol on electrophoretic gel |
| CN101113983A (en) * | 2007-08-30 | 2008-01-30 | 温州医学院 | Application of Palmatine and its derivatives in protein fluorescent detecting |
| CN102053030A (en) * | 2009-11-03 | 2011-05-11 | 广东暨大基因药物工程研究中心有限公司 | Mass spectrum-compatible protein argentation |
| CN201803885U (en) * | 2010-09-13 | 2011-04-20 | 深圳市卫武光明生物制品有限公司 | Gel dyeing and decolorizing device |
| CN102061632A (en) * | 2010-11-19 | 2011-05-18 | 南京工业大学 | Dyeing process of natural organic dye extracted from safflower |
| CN102590099A (en) * | 2011-01-18 | 2012-07-18 | 温州医学院 | Application of 8-phenylamino-1-naphthalene sulfonic acid and derivative thereof to protein fluorescence detection |
| CN102200526A (en) * | 2011-03-15 | 2011-09-28 | 六安初创人乳库有限公司 | Qualitative detection method capable of distinguishing cow milk doped in human milk |
| CN102607920A (en) * | 2012-03-15 | 2012-07-25 | 中国热带农业科学院热带生物技术研究所 | SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) coomassie brilliant blue R250 fast staining solution, staining method and application |
| CN102911189A (en) * | 2012-10-16 | 2013-02-06 | 桂林理工大学 | Salicylaldehyde azine schiff base transition metal compound and application thereof |
| CN103149262A (en) * | 2013-01-11 | 2013-06-12 | 安徽中医学院第一附属医院 | Xinfeng capsule water-soluble protein molecular weight detecting method |
| CN103267666A (en) * | 2013-02-21 | 2013-08-28 | 山东理工大学 | Protein and gamma-polyglutamic acid simultaneous coloring counterstaining method |
| CN103233060A (en) * | 2013-04-26 | 2013-08-07 | 天津科技大学 | Method for analyzing protein expression during fungal fermentation process |
Non-Patent Citations (19)
| Title |
|---|
| A Fluorescence Emission, FT-IR and UV-VIS Absorption Study of the Some Uranium (VI) Schiff Bases Complexes;Th. Măluţan et.al;《Journal of Fluorescence》;Springer;20080219;第18卷;707-713 * |
| Fluorescence staining of salicylaldehyde azine, and applications in the determination of potassium tertbutoxide;Jinlai Yang et.al;《RSC Advances》;RSC;20160318;第6卷;30636-30641 * |
| Fluorescent staining of protein in SDS polyacrylamide gels by salicylaldehyde azine;Mao-Wei Ni et.al;《Electrophoresis》;Wiley;20131025;第34卷;3171-3179 * |
| Organic Crystalline Solids Response to Piezo/thermo Stimulus:Donor-Acceptor (D-A) Attached Salicylaldehyde Azine Derivatives;Xiaotong Chen et.al;《The Journal of Physical Chemistry C》;ACS;20110706;第115卷;14353-14359 * |
| Salicylaldehyde Azines as Fluorophores of Aggregation-Induced Emission Enhancement Characteristics;Weixin Tang et.al;《Journal of Organic Chemistry》;ACS;20090129;第74卷;2163-2166 * |
| SDS-PAGE电泳技术分析蛋白质的研究;高艳利等;《辽宁化工》;20070731;第36卷(第7期);460-463 * |
| Sensitive fluorescent staining for proteomic analysis of proteins in 1-D and 2-D SDSPAGE and its comparison with SYPRO Ruby by PMF;Wei-Tao Cong et.al;《Electrophoresis》;Wiley;20081231;第29卷;摘要，第2节全文 * |
| Spectroscopic and photophysical properties of salicylaldehyde azine (SAA) as a photochromic Schiff base suitable for heterogeneous studies;Marcin Ziółek et.al;《Chemical Physics Letters》;20080919;第464卷;182 * |
| SYPRO Orange and SYPRO Red Protein Gel Stains:One-Step Fluorescent Staining of Denaturing Gels for Detection of Nanogram Levels of Protein;Thomas H. Steinberg et.al;《ANALYTICAL BIOCHEMISTRY》;19961231;第239卷;223-237 * |
| 凝胶上水杨醛吖嗪蛋白质荧光染色技术研究;张慧;《温州医科大学硕士学位论文》;20160330;全文 * |
| 凝胶上蛋白质染色方法研究进展;周旋等;《中国医药生物技术》;20111031;第6卷(第5期);378-381 * |
| 几种可见染色法对特殊蛋白的检测;张美玲等;《温州医学院学报》;20090731;第39卷(第4期);364-366 * |
| 发芽糙米中谷蛋白亚基电泳分析;郭晓娜等;《根食与油脂》;20031231(第11期);19-21 * |
| 在聚丙烯酰胺凝胶上蛋白质、DNA染色方法的研究;朱忠欣;《》;万方医学;20101222;全文 * |
| 应用蛋白质组分析法初步探讨胆囊收缩素在皮质神经元中的信号转导途径;李娟等;《泸州医学院学报》;20061231;第29卷(第5期);420-422 * |
| 改良聚酰胺吸附-高效毛细管电泳内标法测定饮料中的亮蓝和苋菜红;闫正等;《色谱》;20101231;第28卷(第12期);1185-1188 * |
| 聚丙烯酞胺凝胶的蛋白质荧光染色法;梁念慈等;《生物化学与生物物理进展》;19861231(第4期);1页 * |
| 荧光染料结构与性能关系的研究进展;赵德丰等;《染料工业》;19971231;第34卷(第3期);1-6 * |
| 马槟榔甜味蛋白的研究IV·稳定性和变性;丁鸣等;《云南植物研究》;19861231;第8卷(第2期);181-192 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104422659A (en) | 2015-03-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| NAYYAR et al. | Studies in histochemistry XXXI. A method for the determination of protein in millimicrogram quantities | |
| DK343679A (en) | IMMUNOLOGICAL PROCEDURE FOR DETERMINING AN IMMUNOLOGY COMPLEX IN A FLUID AND THE DIAGNOSTIC AGENT USED IN THE PROCEDURE | |
| CN104655846A (en) | Enzyme linked immunosorbent assay kit for detecting progesterone and detection method thereof | |
| Goldring | Protein quantification methods to determine protein concentration prior to electrophoresis | |
| CN105974110A (en) | Immune lateral chromatographic detection system as well as preparation method and application thereof | |
| CN101526525B (en) | Enzyme-linked immunosorbent assay kit suitable for pentachlorophenol residual analysis | |
| CN104422659B (en) | The application of salicylide azine and its derivative in protein fluorescence detection | |
| CN103901199A (en) | Preparation of ELISA kit for detecting plasticizer (DBP) | |
| CN102590099B (en) | The application in protein fluorescence detection of 8-anilino-1-naphthalene sulfonic acid and derivant thereof | |
| CN106950369A (en) | A kind of pig epidemic diarrhea novel antibodies ELISA detection kit | |
| CN105319368A (en) | Enzyme linked immunosorbent assay kit used for detecting zearalenone, and detection method thereof | |
| CN103558372B (en) | Preparation method of antigen standard substance and sample diluting solution for ELISA (enzyme-linked immuno sorbent assay) | |
| JPWO2006132030A1 (en) | Novel compound, reagent for analysis of peptide or protein containing the compound, and analysis method using the analysis reagent | |
| CN101113984A (en) | Application of berberine and its derivates in protein fluorescent detecting | |
| CA1133811A (en) | Diagnostic agent for the proof of urobilinogen | |
| CN105301244B (en) | Detect enzyme linked immunological kit and its application of acid orange | |
| Fischer et al. | Fixation and permeabilization of cells and tissues | |
| CN101113983A (en) | Application of Palmatine and its derivatives in protein fluorescent detecting | |
| CN103808921A (en) | Enzyme-linked immunosorbent assay kit for detecting residual zilpaterol and use method thereof | |
| CN104483294B (en) | The application of calcium carboxylate and its derivative in phosphorylating protein fluoroscopic examination | |
| CN117347617A (en) | Diluent and application thereof, and method for determining analyte in sample | |
| CN202903791U (en) | T-2 toxin ELISA (enzyme-linked immunoabsorbent assay) detection reagent kit | |
| CN101975857B (en) | Virus disaggregating agent and method for disaggregating virus antigen-antibody complex and detecting HCV (Hepatitis C Virus) antigen | |
| CN105319367A (en) | Preparation of enzyme linked immunosorbent assay kit used for detecting plasticizer | |
| US3201202A (en) | Diagnostic procedure |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20221024 Address after: Room 517, 518, 523, 524, 525, Building D, No. 100, Luodongbei Street, Yongzhong Street, Longwan District, Wenzhou City, Zhejiang Province, 325000 Patentee after: Zhejiang Eye Spring Biotechnology Co.,Ltd. Address before: 325035 Room 218, Building C, Community 10, Wenzhou Hi tech Industrial Park, Zhejiang Province Patentee before: WENZHOU UNDERSUN BIOTECHNOLOGY Co.,Ltd. |
|
| TR01 | Transfer of patent right |