Which HRP substrate.
m.davies at herts.ac.uk
Thu Jan 26 06:35:07 EST 2006
I need some help trying to find a Horseradish Peroxidase substrate that
fulfils certain requirements.
1. Soluble in aqueous solution buffered at pH 7. =96 necessary=20
2. Concentration measured by absorbance, not fluorescence, at visible
wavelengths. =96 necessary=20
3. Time stable substrate =96 necessary=20
4. Time stable product =96 needs to be reasonably stable as the
measurements need to be made over the course of at least 4 hours.=20
5. Light stable. - Not necessary, but would be extremely useful.=20
6. Room temperature stable. =96 necessary=20
7. Known chemistry =96 preferable so I can at least understand how the
8. Single stage reaction =96 necessary,=20
As an example I am currently working with TMB. The reaction produces a
product that is then generally stabilised and changed by quenching with
acid. I do not have the option in the final application to add a =
solution after the initial reaction has taken place. I can detect the
presence of the initial product, so its not too bad, but TMB is not
horrendously soluble at pH 7 and is not light stable. The measured =
reaction product, ie before quenching, is a charge transfer complex. I =
not too happy using this, because factors such as dilution change the
response and I am not certain that the product I am measuring is stable
enough for my purposes.
I am not a biologist or even a biochemist. I am a chemist trying to use =
as part of a system to quantitatively detect the formation of hydrogen
Any help is much appreciated.
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