- Intended Use
- BUF066A is a high performance TMB (3,3´, 5, 5´- tetramethylbenzidine) solution, recommended for use in ELISA as a substrate for horseradish peroxidase (HRP).
BUF066A 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.
BUF066A produces a deep blue colour during the enzymatic degradation of H2O2 by horseradish Peroxidase. The reaction may be stopped with 0.25M Sulphuric acid, resulting in a yellow colour read at 450nm.
BUF066A is more sensitive than other TMB reagents offered by Bio-Rad.
- Product Form
- Ready to use TMB solution - liquid
- Store at +4oC. 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).
- Guaranteed until date of expiry. Please see product label.
- For research purposes only
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Applications of TMB Sensitive
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
- Instructions For Use
- 1. It is recommended that 100ul of BUF066A 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 of substrate solution per microtiter well.
3. Allow development of the substrate solution. Development time is typically 5-15 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 0.25M 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 BUF066A is not recommended).
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