Hi everyone,
last time I was doing an ELISA without exposing it to the light and before I added the substrate it was turning blue. Do you have any idea or suggestion of a better substrate?, the substrate I use is one from BD.
thanks a lot!
ABTS (Fisher) is more stable and reliable than TMB. It is a water-soluble peroxidase substrate that yields a measurable green end product for use in ELISA methods. It has lower sensitivity compared to OPD or TMB, less readily oxidized and has a slower color development. It is especially useful if some background results are observed due to higher sensitivity of the other substrates mentioned.
Is your pipette tip contaminated with an HRP containing solution?
Di you pour the TMB into a container that previously held the diluted conjugate for your ELISA? It only takes a trace amount of HRP to turn the TMB solution blue.
It may be Caroline, we use some boxes for incubating wb membrane and I also have use them to prepare the substrate. Thank you for making ne botice this issue. Best, carolina
Hello, I use the one from R&D systems, it works well, no blue color without incubation.
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