TMB 与HRP产生显色反应，在ELISA时，显色剂中的主要试剂是TMB，但现在一般都是分A、B液的，用时很不方面，能不能做成一种试剂并且可以保存够长时间的方法呢？

好象不可以，显色液不太稳定，最好新鲜配置．

引用:

原帖由 pei 于 07-11-9 00:36 发表
TMB 与HRP产生显色反应，在ELISA时，显色剂中的主要试剂是TMB，但现在一般都是分A、B液的，用时很不方面，能不能做成一种试剂并且可以保存够长时间的方法呢？ ...

KPL公司就生产一种试剂的显色液，我也很想知道KPL公司是怎么做到的。

国内 天健生物 有生产单组分TMB底物.  据用户反映还行. 你可以试着联系一下.
(呵呵,其实就是联系我们. 我是天健生物的客服.
我们的TMB很不错,北京上海武汉深圳等地的诊断试剂厂商都有用. :-)
免费电话:800-818-0100.  邮箱:sales at tj-bio.com  网站http://www.tj-bio.com)

为以示公允, 我可以告诉你, 国内做单组分TMB的, 还有北京天根.

[ 本帖最后由 istu 于 07-12-26 09:27 编辑 ]

1-Component TMB Peroxidase Substrate

Catalog: 610-0100, 610-0500, 610-1000
  
610-0100, 1-Component TMB Peroxidase Substrate, 100 ml  
610-0500, 1-Component TMB Peroxidase Substrate, 500 ml
610-1000, 1-Component TMB Peroxidase Substrate, 1000 ml
  
Description
TMB Microwell Peroxidase Substrate (1-Component) develops a deep blue soluble product when reacted with horseradish peroxidase labeled conjugates in microwell plates or other reaction systems. It’s suitable for qualitative or quantitative enzyme immunoassays. It is not suitable for membrane or immunohistochemical staining.

Appearance and Background

The expected appearance of the solution is colorless or light blue. The background of unreacted substrate at 650 nm is < 0.04 OD

Form/Storage/Stability

1-component liquid. No organic solvent.  Store at 2 - 8°C.  Do not freeze. Stable for a minimumof 18 months from date of manufacture with proper storage condition.

Recommended Use

Pour out the desired amount into a clean container and warm to room temperature before use. Solution is ready to use.
Substrate Development: Following incubation with peroxidase labeled conjugate, wash plate thoroughly. Add
100-150 μL substrate solution to each microwell. As the color develops, tap gently to mix. Incubation times will 15-30 minutes, depending on your assay.
To Stop Reaction: For optimal performance, stop reaction by adding equal volume of 1N HCl or 2.0 N H2SO4 to the microwell plate. This will halt color development and will turn the substrate yellow.
To Read Reaction: After stopping, read at a wavelength of 450
nm. Stopped reaction should be read within 30 minutes.





Note:

        To Reduce Substrate Intensity: High background and/or precipitation in the microwells indicate the sign of over-reaction with TMB substrate. To avoid the interferences of substrate precipitation, try to read the results as soon as possible after adding stopping solution. Alternatively, further dilute the primary antibody and/or the HRP conjugate. To reduce the non-specific background reaction, further dilution of the primary antibody and/or HRP labeled conjugate is recommended. Dilution of the substrate is not a recommended option.

        Kinetic Assays:  TMB  Peroxidase Substrate (1-Component) produces a blue color upon reaction with peroxidase. Read at a wavelength of 620 - 650 nm right after addition the substrate.


References

1.        Porstman, T., and Kiessig, S.T. (1992). J. Immunol. Methods 150: 521.
2.        Joseph, P.D., et al (1982),  J. of Biological Chemistry 257:36694675