TMB Accessory Reagent

Ready To Use
Ready to use TMB solution - liquid

Store at 4 degree C. DO NOT FREEZE. This product is photosensitive and should be protected from light. Avoid exposure to heat and contamination with metal ions or peroxidase. Store in bottles made of High Density Polyethylene (HDPE).
Shelf Life: 48 months from date of despatch.

This item is a high performance TMB (3,3´, 5, 5´- tetramethylbenzidine) solution, recommended for use in ELISA as a substrate for horseradish peroxidase (HRP).

This item contains TMB, substrate buffer and hydrogen peroxide in a safe, ready to use solution. The activity of TMB has been optimised to enable increased sensitivity, minimal background and rapid development.

This item produces a deep blue colour during the enzymatic degradation of H2O2 by horseradish Peroxidase. The reaction may be stopped with 0.2M sulphuric acid, resulting in a yellow colour read at 450nm.
Instructions For Use||1. It is recommended that 100ul of this item TMB substrate is used per microtiter well. Pour the desired amount of substrate into a sealed container and allow it to reach room termperature in the dark.
2. Add 100ul substrate solution per microtiter well.
3. Allow development of the substrate solution. Time of development is typically 5-30 minutes. For best results, the plate should be kept in the dark during incubation e.g. wrapped in tinfoil.
4. For kinetic assays, read absorbance at 655nm (blue). For endpoint assays, add an equal volume of 0.2M sulphuric acid and read the absorbance at 450nm (yellow). This endpoint solution is stable for up to one hour.

N.B. If reduced intensity is required, it is recommended that the development time is reduced or the antibody/conjugate is diluted further. (Dilution of This item is not recommended).