BUF055A is a high performance TMB (3,3´, 5, 5´- tetramethylbenzidine) solution, recommended for use in ELISA detection systems as a substrate for horseradish peroxidase (HRP). TMB SIGNAL+ PRESTAINED is stained with a red indicator to eliminate pipetting errors. The red indicator does not interfere with the sensitivity or the stability of the substrate. It does not absorb light at the wavelengths used for the substrate and will disappear after addition of the stop solution.
BUF055A contains TMB, substrate buffer and hydrogen peroxide in a safe, ready to use solution. TMB SIGNAL+ PRESTAINED has been optimised to enable increased sensitivity whilst maintaining increased stability. BUF055A provides minimal background and rapid development.
BUF055A produces a deep blue colour during the enzymatic degradation of H2O2by horseradish peroxidase. For kinetic assays the absorbance is read at 655nm (blue). However, for endpoint assays 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
24 months from date of despatch.
Instructions For Use
1. Pour the desired amount of BUF055A into a sealable container and allow it to reach room temperature in the dark. Note that 0.1ml will be required per well.
2. Add 0.1ml of BUF055A per microtiter well.
3. Allow development of BUF055A. Development time is typically 5-30 minutes. For best results, the plate should be kept in the dark during incubation. i.e. 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.
5. If reduced intensity is required, it is recommended that the development time is reduced or the antibody/conjugate is diluted further. (Dilution of BUF055A is not recommended).
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