Product datasheet
ADHP HRP Substrate Kit ab205810
1 Image
Overview
Product name
ADHP HRP Substrate Kit
Detection method
Fluorescent
Assay time
0h 15m
Assay duration
Multiple steps standard assay
Product overview
ADHP HRP kit (ab205810). The HRP substrate is a combination of 10-Acetyl-3,7dihydroxyphenoxazine (ADHP), a sensitive substrate for HRP, and ADHP Dilution Buffer, a
stabilized H2O2 solution. In the presence of H2O2, HRP converts ADHP to the fluorogenic
product, resorufin. Resorufin is a highly fluorescent molecule, and should be used with excitation
filters in the range of 530 - 540 nm, with bandwidths ≤ 30 nm. The emission filter should be in the
range of 590 - 600 nm, with bandwidths ≤ 30 nm.
Notes
The signal in the wells should be developed for around 10 minutes. Best results will be obtained
if the microplates are developed in the dark, e.g. by covering the microplate with foil. The Stop
Solution provided can be used for stopping HRP-mediated conversion of ADHP to resorufin.
Once added to the wells, signal in the wells is stable for 60 minutes. However, ADHP remaining
in the wells is still light-sensitive, and should be handled under low-light conditions until the
microplate is read.
Tested applications
ELISA
Platform
Microplate (12 x 8 well strips)
Properties
Storage instructions
Store at +4°C. Please refer to protocols.
Components
1 x 96 tests
ADHP (100X)
1 x 120µl
ADHP Dilution Buffer
1 x 15ml
Stop Solution
1 x 2ml
Applications
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Our Abpromise guarantee covers the use of ab205810 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Abreviews
ELISA
Notes
Use at an assay dependent concentration.
ADHP HRP Substrate Kit images
Comparison of ADHP fluorescence (A) and
TMB colorimetric (B) assay readouts. Using
an ELISA based format, standard curves
were generated for a test analyte with
concentrations ranging from 40 fM up to 10
nM. After washing away unbound reactants,
bound HRP was reacted with ADHP or TMB
substrate for 10 minutes before stopping
each reaction with its respective stop solution.
Fluorescence was measured at 540/590
using a FlexStation II384 (Molecular Devices)
ab205810
and absorbance was measured at 450 nm
using an EnVision 2102 Multilabel Reader
(Perkin Elmer).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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Terms and conditions
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