Which HRP substrate.

Matthew Davies m.davies at herts.ac.uk
Thu Jan 26 06:35:07 EST 2006


=91Scuse all

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I need some help trying to find a Horseradish Peroxidase substrate that
fulfils certain requirements.

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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
reaction proceeds.=20
8.	Single stage reaction =96 necessary,=20

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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 =
quenching
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 =
initial
reaction product, ie before quenching, is a charge transfer complex.  I =
am
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.

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I am not a biologist or even a biochemist.  I am a chemist trying to use =
HRP
as part of a system to quantitatively detect the formation of hydrogen
peroxide.

Any help is much appreciated.

Thankyou


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