TMB ELISA Substrate

Product Overview

Description

ProteinFind^TM Anti-beta-Tubulin

Mouse Monoclonal Antibody

TMB ELISA Substrate is a ready-to-use chromogenic substrate for detection of horseradish peroxidase (HRP) activity. HRP can catalyze 3,3',5,5'-tetramethylbenzidine (TMB) to yield a blue color, the maximal absorbance is at 370 nm or 620-652 nm; however, upon addition of the stop solution, the solution turns to yellow and can be measured at 450 nm.

 

Cat. No.	HE101
Specificaation	HC101-01	100 ml
Storage:	at 2-8 o C in dark for one year
Notes	If shorter reaction time is needed, incubate the solution at 37 o C incubator in dark after adding TMB Solution.

 

Procedures
1. In ELISA experiment, after incubation with HRP-conjugated secondary antibody, wash ELISA plate 3

    times with PBST (PBS containing 0.05% Tween-20), 5 minutes each time.
2. Add 100 µl of TMB Solution (equilibrated to room temperature in advance) to each well. Incubate for

    3-30 minutes or longer time in dark until the solution changes to expected color.
3. (optional) Add 100 µl of stop solution to each well (prepared by user, 0.5 M H 2 SO 4 or 1 M HCl).
4. Measure the absorbance at 370 nm or 620-650 nm (the reaction has not been stopped), or measure the

    absorbance at 450 nm (the reaction with stop solution).

 

Packaging