(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.
TMB is a one component system that is ready to use from the bottle.
Store at 4oC.
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