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3,3′,5,5′-Tetramethylbenzidine (TMB) Substrate, ELISA

Cat No. GTX14696

Application

ELISA
Package
500 ml ($319),
100 ml ($79)
PRODUCT

Summary

The GTX14696 is supplied as a ready‐to‐use solution. Peroxidase reacts with the substrate solution to produce a soluble blue reaction product that can be read at 370 nm or 620‐650 nm for ELISA applications. Stop solutions can be added to prevent further color development and reading the absorbance values at the desired wavelengths.
A soluble yellow product develops after stopping and stabilizing with a 450 nm stop solution.
The developing soluble blue product can be stopped and stabilized by using a 650 nm stop solution.

APPLICATION

Application Note

Allow the substrate to equilibrate at room temperature (~25ºC) prior to use.
End-Point Assays:
(1) Add 100 μL of equilibrated substrate solution to each microwell.
(2) Allow the substrate reaction to develop a blue soluble reaction by incubating for the optimal time determined for the assay.
(3) The reaction should be stopped by using a 450 nm or 650 nm stop solution.
(4) Measure the absorbance at 450 nm or 650 nm of each microwell within 1 hour.
Kinetic Assays:
(1) Add 100 μL of equilibrated substrate solution to each microtiter plate well.
(2) Measure the absorbance at 650 nm at multiple time points.
(3) Calculate the rate of absorbance change for each microwell.

PROPERTIES

Form

Liquid

Storage

Stable for 2.5 years at 4ºC. Protect from light.

Note

For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.

TARGET

Background

GeneTex TMB substrate is supplied as a one-component- ready-to-use solution. It is colorless or very light yellow in appearance. When TMB reacts with peroxidase, a blue liquid is yielded (oxidized product). The reaction can be terminated using 0.6N HCl. GeneTex TMB substrate is not recommended for membrane or immunohistochemistry applications where a colored precipitate is desired.

REFERENCE

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REVIEW

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Package List Price ($)
$ 319
$ 79