CN102866249B - Tetramethylbenzidine (TMB) developing system - Google Patents
Tetramethylbenzidine (TMB) developing system Download PDFInfo
- Publication number
- CN102866249B CN102866249B CN201210335101.5A CN201210335101A CN102866249B CN 102866249 B CN102866249 B CN 102866249B CN 201210335101 A CN201210335101 A CN 201210335101A CN 102866249 B CN102866249 B CN 102866249B
- Authority
- CN
- China
- Prior art keywords
- tmb
- damping fluid
- tmb substrate
- substrate damping
- prepared
- 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.)
- Expired - Fee Related
Links
Landscapes
- Liquid Developers In Electrophotography (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a tetramethylbenzidine (TMB) developing system, which comprises TMB storage liquid and TMB substrate buffering liquid mother liquid, wherein the TMB storage liquid is prepared by the following raw materials by percent weight volume: 4,5 to 5.2g TMB, 450ml dimethylsulfoxide (DMSO) and 50ml glycerol; and the TMB substrate buffering liquid mother liquid is prepared by the following raw materials by percent weight volume: 100 to 120g sodium acetate. 3H2O, 90 to 110g citric acid.H2O and 800mL distilled water, and the PH value is adjusted by thick hydrochloric acid to 3.0 to 4.0; and placing the distilled water to fix the volume to 1000mL to prepare the TMB substrate buffering liquid mother liquid. A developing agent is prepared by the following steps of (1) preparing TMB substrate buffering liquid working liquid: 10mL TMB substrate buffering liquid mother liquid, 0.30 to 0.40mL hydrogen peroxide ( H2O20 (30 percent); and adding the distilled water to fix the volume to 1000mL to prepare the TMB substrate buffering liquid working liquid; and (2) mixing the TMB storage liquid and the TMB substrate buffering liquid working liquid according to a volume ratio of 20 to 100: 1000 to prepare the TMB developing agent.
Description
Technical field
The present invention relates in vitro diagnostic reagent, specifically TMB color development system.
Background technology
Setting up enzyme linked immunosorbent detection ELISA (Enzyme-Linked-Immuno-Sorbent Assays) technology platform of efficient stable, is many laboratories and clinical examination worker long-cherished wish for many years.In recent years, due to the widespread use that ELISA method detects in clinical detection and bio-pharmaceutical, testing requirement develops into quantitative test today from qualitative detection before.Along with the conversion of this testing requirement, requirements at the higher level are proposed for the accuracy of detection method and stability.The quality of data detected also is developed into by " almost " before " can not be poor " of today.In order to ensure the reliable and stable of ELISA method, need optimization and the close fit of a lot of link, because the quality of colour developing has directly had influence on the success or failure of ELISA method and the quality of data of detection, therefore, although the preparation of TMB developer is the final step of the method, is a step of most critical.
TMB and tetramethyl benzidine (3,3 ', 5,5 '-Tetramethylbenzidine) are a kind of fat-soluble stronger materials, through with HRP effect after product aobvious blue, visual comparison is distinct.TMB character is more stable, can wiring solution-forming reagent, only need and H
2o
2solution mixed application liquid, directly can make substrate and use.In addition, TMB has the advantages such as non-carcinogenesis again, therefore applies increasingly extensive in ELISA.With HCL or H after reaction
2sO
4after termination, TMB product is by blueness in yellow, and use microplate reader to detect, the suitableeest absorbing wavelength is 450nm.
Current TMB developer compound method is a lot, and commercially available developer is not only expensive, and it is short to store the term of validity, in use high to the accuracy requirement of operation; Have a lot of laboratory to prepare voluntarily, but compound method is varied does not have unified standards and norms; The TMB developer used in research paper is also multifarious, for detect data to when checking cause difficulty.Therefore need to develop for the sensitive efficient, stable, easy-to-use of in vitro diagnosis and to configure simple TMB color development system be the task of top priority.
Summary of the invention
Object of the present invention be exactly to provide a kind of be suitable for laboratory use sensitive efficient, stable, easy-to-use, configure enzyme linked immunological TMB color development system simple and with low cost.
The object of the present invention is achieved like this:
This TMB color development system, comprises TMB storage liquid and tmb substrate damping fluid mother liquor, and wherein, described TMB storage liquid is prepared by following w/v by following raw material:
TMB 4.5～5.2g
DMSO 450mL
Glycerine 50mL;
Described tmb substrate damping fluid mother liquor is prepared by following w/v by following raw material:
Sodium acetate 3H
2o 100 ~ 120g
Citric acid H
2o 90 ~ 110g
Add distilled water 800mL
PH to 3.0 ~ 4.0 are regulated with concentrated hydrochloric acid
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid mother liquor.
In TMB color development system of the present invention, described TMB storage liquid is preferably prepared by following w/v by following raw material:
TMB 5.0g
DMSO 450mL
Glycerine 50mL;
Described tmb substrate damping fluid mother liquor is preferably prepared by following w/v by following raw material:
Sodium acetate 3H
2o 113.0g
Citric acid H
2o 105.0g
Add distilled water 800mL
PH to 3.6 is regulated with concentrated hydrochloric acid
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid mother liquor.
TMB color development system of the present invention, can be made into developer by the following method:
1. tmb substrate damping fluid working fluid is prepared
Tmb substrate damping fluid mother liquor 10mL
H
2O
2（30%） 0.30～0.40mL；
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid working fluid;
2. described TMB storage liquid and described tmb substrate damping fluid working fluid are the obtained TMB developer of ratio mixing of 20 ~ 100: 1000 by volume.
TMB color development system of the present invention, during tmb substrate damping fluid working fluid described in preparation, H
2o
2(30%) addition is preferably 0.33mL.TMB storage liquid and tmb substrate damping fluid working fluid are preferably the obtained TMB developer of ratio mixing of 1: 50 by volume.
The compound method of TMB developer comprises the following steps:
1. TMB storage liquid is prepared by following raw material by following w/v:
TMB 5.0g
DMSO 450mL
Glycerine 50mL;
2. tmb substrate damping fluid mother liquor is prepared by following raw material by following w/v:
Sodium acetate 3H
2o 100 ~ 120g
Citric acid H
2o 90 ~ 110g
Add distilled water 800mL
PH to 3.0 ~ 4.0 are regulated with concentrated hydrochloric acid
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid mother liquor;
3. tmb substrate damping fluid working fluid is prepared
Tmb substrate damping fluid mother liquor 10mL
H
2O
2（30%） 0.30～0.40mL；
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid working fluid;
4. described TMB storage liquid and tmb substrate damping fluid working fluid are the obtained TMB developer of ratio mixing of 20 ~ 100: 1000 by volume.
The compound method of TMB developer is preferably prepared according to the following steps:
1. TMB storage liquid is prepared by following raw material by following w/v:
TMB 5.0g
DMSO 450mL
Glycerine 50mL;
2. tmb substrate damping fluid mother liquor is prepared by following raw material by following w/v:
Sodium acetate 3H
2o 113.0g
Citric acid H
2o 105.0g
Add distilled water 800mL
PH to 3.6 is regulated with concentrated hydrochloric acid
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid mother liquor.
3. tmb substrate damping fluid working fluid is prepared
Tmb substrate damping fluid mother liquor 10mL
H
2O
2（30%） 0.33mL；
Add distilled water to be settled to 1000mL and to obtain tmb substrate damping fluid working fluid;
4. described TMB storage liquid and tmb substrate damping fluid working fluid are the obtained TMB developer of ratio mixing of 1: 50 by volume.
In the present invention, described TMB storage liquid is mixed to dissolve by TMB, DMSO and glycerine to obtain.First the TMB taken is joined in DMSO and stir and evenly mix, dissolve completely to TMB, then add the glycerine measured and stir and evenly mix.
This TMB storage liquid keeps in Dark Place in 4 DEG C, can term of life be 3 years
In the present invention, the process for preparation of described tmb substrate damping fluid mother liquor: by the sodium acetate 3H taken
2o, citric acid H
2o in addition distilled water is stirred to salinity and dissolves completely, and regulate PH to 3.0 ~ 4.0 with concentrated hydrochloric acid, preferred PH is 3.6, then makes through constant volume.
This tmb substrate damping fluid mother liquor keeps in Dark Place in 4 DEG C, can term of life be 3 years.
TMB color development system of the present invention is two component color development system, and it should be prepared in use at any time, should use after having prepared in 30 minutes.Tmb substrate damping fluid mother liquor, when preparing, is first become tmb substrate damping fluid working fluid, then presses configuration proportion by color development system of the present invention, obtained TMB developer.
Accompanying drawing illustrates:
Fig. 1 is proportioning and the Properties of Organic Reagents graph of a relation of the made TMB storage liquid of embodiment 1 and tmb substrate damping fluid working fluid.
Embodiment
Embodiment 1
(1) take off by consumption and state raw material preparation TMB storage liquid:
TMB 5g
DMSO 450mL
Glycerine 50mL.
Made TMB storage liquid keeps in Dark Place in 4 DEG C, the term of validity 3 years, ready access upon use.
(2) take off by consumption and state raw material preparation tmb substrate damping fluid mother liquor:
Sodium acetate 3H
2o 113.0g
Citric acid H
2o 105.0g
Add distilled water 800mL
PH to 3.6 is regulated with concentrated hydrochloric acid
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid mother liquor.
Made tmb substrate damping fluid mother liquor keeps in Dark Place in 4 DEG C, the term of validity 3 years, ready access upon use.
(3) TMB developer is prepared:
1. tmb substrate damping fluid working fluid is prepared:
Tmb substrate damping fluid mother liquor 10mL
H
2O
2（30%） 0.33mL；
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid working fluid.
2. get and add 0.2mL TMB storage liquid in 10mL tmb substrate damping fluid working fluid and obtain TMB developer.
The TMB developer prepared should use in 30 minutes.
The compound method of embodiment 2-10 is with embodiment 1, and the pulp furnish of each embodiment carries out according to table 1.
In table 1 embodiment 2-10, in TMB color development system, various related component contains scale
Test one, TMB color development system color developing of the present invention test
Example 1 to 10 made TMB developer carries out color developing test by following method of testing, and institute's test data is as shown in table 2.As can be seen from this test figure, the made TMB developer of embodiment 1 to 10 all can meet color developing requirement.
TMB chromogenic reagent performance test methods:
1) being diluted with coating buffer 1:10000 by human IgG (Sigma I-4506) is standard S
1(5 μ g/mL).
2) by human IgG coating buffer doubling dilution 6 concentration gradient: S
2(2.5 μ g/mL), S
3(1.25 μ g/mL), S
4(0.63 μ g/mL), S
5(0.31 μ g/mL), S
6(0.16 μ g/mL) and S
7(0.08 μ g/mL).Blank B is coating buffer (not adding human IgG).
3) above-mentioned standard solution 100 μ L is added in corresponding hole, 4 DEG C of night incubation.
4) to turn liquid pat dry residual liquid.
5) 3 times are cleaned with 300 μ L cleansing solutions.
6) add 300 μ L confining liquids in each hole, hatch 1h for 37 DEG C.
7) to turn liquid pat dry residual liquid.
8) with confining liquid by goat anti-human igg-HRP(Sigma A-6178) dilute 8000 times, add 100 μ L in each hole, hatch 1h for 37 DEG C.
9) to turn liquid pat dry residual liquid, clean 5 times with cleansing solution.
10) 100 μ l are added, TMB nitrite ion in every hole, colour developing 15min.
11) stop buffer is added: H
2sO
4(2M) 50 μ L, microplate reader reading, according to OD
450 nmcalculate.
12) data Microplate Manager PC software analysis process.
Wherein:
Coating buffer: Na
2cO
315 mM, NaHCO
335 mM, NaCl 0.2 M;
Confining liquid: 1%BSA resists for diluting two, avoids non-specific binding;
TBS prepares: Tris 12.1g+NaCl 9.0g+H
2o 600ml(is settled to 1000ml), use concentrated hydrochloric acid adjust pH to 7.5.
The preparation (TBST) of cleansing solution: 1000ml TBS+0.5ml Tween-20.
Mentioned reagent is domestic reagent (analyzing pure)
Instrument: ELISA Plate (U.S. Corning Costar 2592), microplate reader (molecule instrument SpectraMax M2 type).
Other are conventional instrument.
(in table, data are OD to table 2 embodiment 1-10 test findings
450nm)
The sensitivity comparison test of test two, the same correlation technique of TMB color development system of the present invention (product)
1, by 2010 editions pharmacopeia formulated nitrite ions:
1. substrate buffer solution (0.005mol/L sodium acetate-citrate buffer) is prepared: take sodium acetate 0.68g, citric acid (C
6h
8o
7h
2o) 1.05g, is dissolved in water and is diluted to 1000ml, adjust pH to 3.6.
2. prepare substrate solution A: take 3,3 ＇ 5,5 ＇-tetramethyl benzidine (TMB) 0.08g, add dimethyl sulfoxide (DMSO) 40ml and dissolves, add methyl alcohol 60ml, mixing, adds substrate buffer solution 100ml, and lucifuge stirs 2 little of dissolving completely, standing 4 hours of room temperature.
3. prepare substrate solution B and measure 1.5% superoxol 3.2ml, add substrate buffer solution to 1000ml.
4. the mixing of substrate solution A, B equal-volume is got before use.
5. stop buffer 2mol/L sulfuric acid solution.
2, get TMB developer, the aforementioned developer by 2010 editions pharmacopeia formulated and commercially available Beijing Products that the present embodiment 1 is prepared, carry out the sensitivity test of TMB developer as follows, obtain test data as shown in table 3.
Wherein, can find out in the following description " when single component TMB peroxidase substrate and horseradish peroxidase connector react; produce a kind of navy blue product; be suitable for quantitatively and the enzyme-linked immuno assay of qualitative analysis " of the instructions of commercially available Beijing Products, in its A B biliquid tmb substrate liquid product, main active principle is TMB, and its usable range for product, use-pattern and application target thereof describe substantially identical with this patent.Be described to again in instructions and " stop: the H adding HCl or 2N of isopyknic 1N
2sO
4carry out cessation reaction.In enzyme mark hole, reactant liquor becomes yellow from blueness.Reading: after cessation reaction, 30 minutes inherent 450nm place readings.", the above, illustrate that the TMB color development system of this commercially available prod and the application has comparability.
The sensitivity comparison test of the same correlation technique of TMB color development system of the present invention (product) is carried out by the chromogenic reagent performance test methods described in test one, wherein, only the 10th) the TMB nitrite ion used in step is different, the operation of other steps is identical, often kind of developer, each detection sample repeats 3 times.
Results contrast is as shown in table 3:
Table 3: (in table, data are OD to the test findings of different formulations developer
450nmmean value ± SD)
As can be seen from Table 3: this color development system
other two kinds of color development system of data fundamental sum
substantially suitable, illustrate that this color development system has higher sensitivity, comparatively other Color Appearance System height about 2 times; And the lower and degree of fitting of the colour developing mxm. of this color development system, blank value all comparatively has advantage both other.
The stability test of test three, TMB color development system and benefit analysis
Described stability test is according to the research technique in test one, prepares developer at every turn and use standard items S by embodiment 1
5test, this test is carried out successively with the prolongation in the storage time of bi-component storage liquid, carry out once at interval of half a year, namely TMB storage liquid and tmb substrate damping fluid mother liquor store half a year, 1 year, a year and a half, 2 years respectively, two-and-a-half years and 3 years time, sampling preparation TMB developer also carries out color developing test and outcome record, logging is as shown in table 4, can check the component of this color development system thus within effective storage life of 3 years, whether the performance of its made developer all can meet request for utilization.
Table 4 is embodiment 1-10
made TMB developer 3 years study on the stability tables of data(in table, data are OD
450nm)
As can be seen from the test findings shown in table 4, the 3 years storage stabilities of the bi-component storage liquid in this color development system are fine, have accumulated 3 years by the study on the stability of above-mentioned test, and developer performance is good.
And by document Investigation, the bi-component storage liquid storage term of validity that result shows other color development system mostly is 1 year or is less than 1 year, be significantly shorter than 3 years of the bi-component storage liquid of color development system of the present invention or the storage term of validity of longer time.
Test four, the test of TMB color development system ease for use and benefit analysis
This test method is with TMB chromogenic reagent performance test methods in test 1, test findings as shown in Figure 1, wherein 1: 1000,2: 1000,5: 1000,10: 1000,20: 1000,40: 1000,80: 1000 and 100: 1000 refer to preparation TMB developer time, the volume ratio of TMB storage liquid and described tmb substrate damping fluid working fluid, TMB storage liquid wherein and described tmb substrate damping fluid working fluid are all with the compound method of embodiment 1.
As can be seen from Figure 1: be 1: 1000,2: 1000,5: 1000,10: 1000 in TMB storage liquid and described tmb substrate damping fluid working fluid volume ratio, namely lower than 20:1000(1:50) time, made TMB developer can not meet colour developing requirement.In TMB storage liquid and described tmb substrate damping fluid working fluid volume ratio at 20:1000(1:50) to 100:1000(1:10) between time, all can meet test colour developing requirement.It can be said that bright, color development system of the present invention, the ratio range of TMB storage liquid and described tmb substrate damping fluid working fluid is fairly widespread, testing requirements can be met in the ratio range of 20 ~ 100:1000, stability and the ease for use of TMB color development system of the present invention can be embodied thus, also illustrate that the durable scope of system of the present invention is larger, thus the operation requirements for user can be reduced.With commercially available prod illustrate with document in fixing component compatibility ratio this point must be used to compare for the many regulations of TMB color development system of two component, this color development system on the proportional range of compatibility with clear superiority not only has positive effect in practical operation, also the stability of the inherence of this color development system is embodied from another aspect, namely color development system itself has stronger stability, can not have influence on final color developing effect in certain scope because of the change of solution.This performance of this color development system in ease for use, also for relevant quality data testing requirement provides prerequisite guarantee.
Consider from the potential danger of saving reagent and minimizing organic solvent, the present invention selects the volume ratio proportioning of TMB storage liquid and described tmb substrate damping fluid working fluid to be 1:50.
Summarize above-mentioned, the TMB color development system that the present invention relates to not only has the sensitive feature such as efficient, stable, easy-to-use, separately, sharpest edges are that the TMB color development system that the present invention relates to is that in existing compound method, component uses minimum, reach test objective and only need 8 kinds of compositions, also looser to the requirement of component, test colour developing requirement can be reached easily, there is the requirement as above describing TMB color development system that the present invention of speciality relates to and be applicable in vitro diagnosis and coherent detection, for the use improving TMB color development system further, there is important guiding effect.
Claims (6)
1. a TMB color development system, comprises TMB storage liquid and tmb substrate damping fluid mother liquor, it is characterized in that,
Described TMB storage liquid is prepared by following w/v by following raw material:
TMB 4.5～5.2g
DMSO 450mL
Glycerine 50mL;
Described tmb substrate damping fluid mother liquor is prepared by following w/v by following raw material:
Sodium acetate 3H
2o 100 ~ 120g
Citric acid H
2o 90 ~ 110g
Add distilled water 800mL
PH to 3.0 ~ 4.0 are regulated with concentrated hydrochloric acid
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid mother liquor.
2. TMB color development system according to claim 1, is characterized in that,
Described TMB storage liquid is prepared by following w/v by following raw material:
TMB 5.0g
DMSO 450mL
Glycerine 50mL;
Described tmb substrate damping fluid mother liquor is prepared by following w/v by following raw material:
Sodium acetate 3H
2o 113.0g
Citric acid H
2o 105.0g
Add distilled water 800mL
PH to 3.6 is regulated with concentrated hydrochloric acid
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid mother liquor.
3. TMB color development system according to claim 1 and 2, is characterized in that, be made into developer by the following method:
1. tmb substrate damping fluid working fluid is prepared
Tmb substrate damping fluid mother liquor 10mL
30% H
2O
2 0.30～0.40mL；
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid working fluid;
2. described TMB storage liquid and described tmb substrate damping fluid working fluid are the obtained TMB developer of ratio mixing of 20 ~ 100: 1000 by volume.
4. TMB color development system according to claim 3, is characterized in that, 1. prepares in tmb substrate damping fluid working fluid and adds 30% H
2o
2for 0.33mL, 2. described TMB storage liquid and tmb substrate damping fluid working fluid are the obtained TMB developer of ratio mixing of 1: 50 by volume.
5. a compound method for TMB developer, is characterized in that comprising the following steps:
1. TMB storage liquid is prepared by following raw material by following w/v:
TMB 5.0g
DMSO 450mL
Glycerine 50mL;
2. tmb substrate damping fluid mother liquor is prepared by following raw material by following w/v:
Sodium acetate 3H
2o 100 ~ 120g
Citric acid H
2o 90 ~ 110g
Add distilled water 800mL
PH to 3.0 ~ 4.0 are regulated with concentrated hydrochloric acid
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid mother liquor;
3. tmb substrate damping fluid working fluid is prepared
Tmb substrate damping fluid mother liquor 10mL
30% H
2O
2 0.30～0.40mL；
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid working fluid;
4. described TMB storage liquid and tmb substrate damping fluid working fluid are the obtained TMB developer of ratio mixing of 20 ~ 100: 1000 by volume.
6. the compound method of TMB developer according to claim 5, is characterized in that comprising the following steps:
1. TMB storage liquid is prepared by following raw material by following w/v:
TMB 5.0g
DMSO 450mL
Glycerine 50mL;
2. tmb substrate damping fluid mother liquor is prepared by following raw material by following w/v:
Sodium acetate 3H
2o 113.0g
Citric acid H
2o 105.0g
Add distilled water 800mL
PH to 3.6 is regulated with concentrated hydrochloric acid
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid mother liquor;
3. tmb substrate damping fluid working fluid is prepared
Tmb substrate damping fluid mother liquor 10mL
30% H
2O
2 0.33mL；
Add distilled water to be settled to 1000 mL and to obtain tmb substrate damping fluid working fluid;
4. described TMB storage liquid and tmb substrate damping fluid working fluid are the obtained TMB developer of ratio mixing of 1: 50 by volume.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210335101.5A CN102866249B (en) | 2012-08-15 | 2012-09-12 | Tetramethylbenzidine (TMB) developing system |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210288205.5 | 2012-08-15 | ||
| CN201210288205 | 2012-08-15 | ||
| CN201210335101.5A CN102866249B (en) | 2012-08-15 | 2012-09-12 | Tetramethylbenzidine (TMB) developing system |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102866249A CN102866249A (en) | 2013-01-09 |
| CN102866249B true CN102866249B (en) | 2015-01-21 |
Family
ID=47445240
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210335101.5A Expired - Fee Related CN102866249B (en) | 2012-08-15 | 2012-09-12 | Tetramethylbenzidine (TMB) developing system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102866249B (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103712982B (en) * | 2013-12-13 | 2016-02-03 | 山东博科生物产业有限公司 | A kind of highly sensitive TMB nitrite ion and preparation method thereof |
| CN103698505B (en) * | 2013-12-13 | 2015-06-10 | 山东博科生物产业有限公司 | Efficient stable TMB (Tetramethylbenzidine) color-developing liquor and preparation method thereof |
| CN103645181B (en) * | 2013-12-13 | 2016-02-17 | 山东博科生物产业有限公司 | A kind of enzyme linked immunological assay chromogenic substrate solution |
| CN103913566B (en) * | 2014-04-11 | 2016-01-06 | 苏州浩欧博生物医药有限公司 | A kind of enzyme linked immunological chromogenic substrate and preparation method thereof |
| CN104459114B (en) * | 2014-11-25 | 2016-03-30 | 成都威尔诺生物科技有限公司 | A kind of anti-interference one-component chromogenic enzyme substrate reagent |
| CN105548173A (en) * | 2016-03-12 | 2016-05-04 | 北京易科拜德科技有限公司 | Adiponectin detection kit |
| CN105974107B (en) * | 2016-07-13 | 2018-01-30 | 广州捷倍斯生物科技有限公司 | One-component TMB nitrite ions and its compound method |
| CN110006881B (en) * | 2019-04-01 | 2020-09-11 | 深圳大学 | Detection kit for iodine content in urine and detection method for iodine content in urine |
| CN113686431B (en) * | 2021-07-30 | 2022-09-20 | 四川大学 | Color development liquid for ultraviolet visual detection and application thereof |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080008760A1 (en) * | 2003-04-14 | 2008-01-10 | Alberto Bianco | Functionalized carbon nanotubes, a process for preparing the same and their use in medicinal chemistry |
| CN100582779C (en) * | 2006-03-28 | 2010-01-20 | 北京大学 | Enzyme-linked immune quantitative detecting method for pavarin and its reagent kit |
| CN102147407A (en) * | 2010-12-31 | 2011-08-10 | 中华人民共和国嘉兴出入境检验检疫局 | Method for detecting parachloroaniline and enzyme linked immuno-absorbent assay kit special for parachloroaniline |
| CN102154527A (en) * | 2011-05-12 | 2011-08-17 | 遵义医学院附属医院 | Method for rapidly detecting multi-drug resistant tuberculosis |
-
2012
- 2012-09-12 CN CN201210335101.5A patent/CN102866249B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN102866249A (en) | 2013-01-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102866249B (en) | Tetramethylbenzidine (TMB) developing system | |
| Armbruster et al. | Limit of blank, limit of detection and limit of quantitation | |
| CN105954266A (en) | Magnetic particle-based quantitative chemiluminescent assay kit for anti-ribosome P protein antibody IgG, and preparation and detection methods thereof | |
| CN103149370B (en) | Lipoprotein (a) detection kit | |
| CN104215770A (en) | Two-particle-based retinol binding protein detection kit | |
| CN104198732A (en) | Kit for detecting content of neutrophil gelatinase-associated lipocalin (NGAL) | |
| CN105954267A (en) | Magnetic particle-based quantitative chemiluminescent assay kit for anti-histone antibody IgG, and preparation and detection methods thereof | |
| CN104897864B (en) | The enzyme linked immunological kit of detection amantadine and application thereof | |
| CN104237513A (en) | Thyroid peroxidase antibody magnetic-particle chemiluminescence immune quantitative testing kit | |
| CN103869070A (en) | Enzyme-linked immunosorbent assay kit for dibutyl phthalate detection, and application thereof | |
| CN104076154A (en) | Enzyme linked immunosorbent assay kit detecting folic acid and application thereof | |
| CN105929166A (en) | Magnetic particle-based quantitative chemiluminescent assay kit for anti-LKM-1 antibody IgG, and preparation and detection methods thereof | |
| CN102798726B (en) | Cobalamin chemiluminescence immunoassay immue quantitative detection reagent box and preparation method thereof | |
| CN109444415A (en) | Metanephrine electrochemiluminescent immunoassay detection kit | |
| CN1912594A (en) | Chemiluminescence investigating method of creatinine in serum | |
| CN101498715B (en) | Anti-cardiolipin antibody detection kit and use thereof | |
| CN105044325A (en) | Enzyme-linked immunosorbent assay kit for detecting triazophos and application of enzyme-linked immunosorbent assay kit | |
| CN103558372B (en) | Preparation method of antigen standard substance and sample diluting solution for ELISA (enzyme-linked immuno sorbent assay) | |
| CN107044977A (en) | A kind of tyrosine phosphatase antibody chemical luminescence immunity detection reagent and preparation method thereof | |
| CN103969445B (en) | The preparation of heme-manganese dioxide composites and the method for detecting human IgG thereof | |
| CN105929172A (en) | Magnetic particle-based quantitative chemiluminescent assay kit for anti-PM-Scl antibody IgG, and preparation and detection methods thereof | |
| Lee et al. | Effect of differences in serum creatinine estimation methodologies on estimated glomerular filtration rate | |
| CN103913566B (en) | A kind of enzyme linked immunological chromogenic substrate and preparation method thereof | |
| ATE524738T1 (en) | IMMUNOASSAY METHOD AND KIT TO BE USED | |
| CN106198959A (en) | Serum hyaluronic acid chemiluminescence immune detection reagent kit and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150121 Termination date: 20200912 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |