ReadiUse™ ABTS Substrate Solution *Optimized for ELISA Assays with HRP Conjugates*

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Chemical structure for ReadiUse™ ABTS Substrate Solution *Optimized for ELISA Assays with HRP Conjugates*
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CAS #30931-67-0
Storage Refrigerated (2-8 °C)
Minimize light exposure
Category Enzyme Detection
Horseradish Peroxidase (HRP)
Horseradish peroxidase (HRP) and HRP conjugates facilitate the ABTS oxidation in the presence of hydrogen peroxide, turning ABTS into its blue-green oxidized product. This chromogenic reaction is widely used for quantify HRP in ELISA assays. The oxidized ABTS product has the absorption maximum of 420 nm that can easily be followed with a spectrophotometer. ReadiUse™ ABTS Solution is optimized for ELISA assays that use HRP or HRP-labeled conjugates and hydrogen peroxide in microwell plates or test tubes. Our ABTS solution allows HRP reaction done in a single addition. The assay solution changes its color to light green upon its reaction with HRP or HRP conjugates in the presence of hydrogen peroxide.


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This protocol only provides a guideline, and should be modified according to your specific needs.
At a glance

Important notes
Warm ReadiUse™ ABTS Solution to room temperature before use. The reagent is to be used as supplied, no dilution is required.

Key parameters
Instrument:Absorbance microplate reader
Absorbance:420 nm
Recommended plate:Solid white
Sample experimental protocol
  1. Wash the assay plate following the incubation of HRP-labeled reagent.

  2. Add 100 µL of ReadiUse™ ABTS Solution into each well.

  3. Incubate the plate at room temperature for 15 – 30 min. Note: The incubation time varies depending on the assay conditions.

  4. Measure the absorbance signal at 415 ± 10 nm (maximum at 420 nm) with an ELISA microplate reader. Note: If desired, the reaction can be stopped by adding an equal volume of 1% SDS or 0.01% sodium azide into 0.1 M citric acid. Stopped reaction should be read within 30 minutes.
Example data analysis and figures

Figure 1. Chemical structure for ReadiUse™ ABTS Substrate Solution *Optimized for ELISA Assays with HRP Conjugates*
AAT Bioquest provides high-quality reagents and materials for research use only. For proper handling of potentially hazardous chemicals, please consult the Safety Data Sheet (SDS) provided for the product. Chemical analysis and/or reverse engineering of any kit or its components is strictly prohibited without written permission from AAT Bioquest. Please call 408-733-1055 or email if you have any questions.

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