BUF042B is a high performance TMB (3,3´, 5, 5´- tetramethylbenzidine) solution, recommended for use in ELISA as a substrate for horseradish peroxidase (HRP).
BUF042B 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.
BUF042B produces a deep blue colour during the enzymatic degradation of H2O2by Horseradish Peroxidase. The reaction may be stopped with 0.2M sulphuric acid, resulting in a yellow colour read at 450nm.
Application
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 visitwww.abdserotec.com/protocols.
Application Name
Yes
No
Not Determined
Suggested Dilution
ELISA
Neat
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.
Storage
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).
Shelf Life
18 months from date of despatch.
Instructions For Use
1. It is recommended that 100ul of BUF042B 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 BUF042B is not recommended).
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