Cell Signaling Technology

Product Pathways - PathScan ELISA

TMB Substrate #7004

Applications Species-Reactivity Sensitivity MW (kDa) Isotype

Species cross-reactivity is determined by western blot.

Applications Key: ELISA=ELISA,


TMB Substrate used is ready to use for ELISA detection. Reaction between the substrate and immobilized horseradish peroxidase (HRP) conjugated secondary antibodies in the ELISA wells produces a blue colored solution. Color intensity and development time will vary depending on assay sensitivity and conditions. After reaching the desired color intensity, the reaction is terminated by addition of an acidic STOP solution which changes the solution color from blue to yellow. While the results will remain stable for one hour following termination, the plate should be analyzed promptly on a microwell reader at 450 nm. TMB is light sensitive and is therefore packaged in amber bottles to protect the solution from direct sunlight. Recommended storage temperature is 4ºC.


Directions for Use

(a) After incubation with horseradish peroxidase (HRP) conjugated secondary antibody, wash plates with PBS/0.05% Tween-20. 
 (b) Add 100 μl of TMB solution/well. Incubate for specific amount of time or until desired signal is achieved. See product datasheet for recommended incubation times for specific antibody pair. 
 (c) Add 100 μl of STOP solution. 
 (d) Read Absorbance at 450 nm.

(a)与辣根过氧化物酶(HRP)结合的二抗一起孵育后,用含有0.05%的Tween-20的PBS洗板。 (b)每孔加入100ulTMB溶液。孵育特定的时间或者直到产生所需要的信号。请参考说明书中建议的孵育时间。 (c)加入100ul终止液 (d)在450nm处读取吸光值


TMB (3,3', 5,5"-tetramethylbenzidine) is used as a substrate to generate detectable signal in ELISA. The reaction between the TMB substrate and a peroxidase, typically horseradish peroxidase (HRP), produces a measureable color change that correlates with analyte level.

TMB( 3,3‘,5,5’-四甲基联苯胺)在ELISA中作为底物以产生可检测光。TMB与过氧化物酶,典型的是辣根过氧化物酶(HRP)之间的反应可以产生可测量的与分析物相关颜色变化。

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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