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ReadiUse™ TMB Substrate Solution *Optimized for ELISA Assays with HRP Conjugates*
Horseradish peroxidase (HRP) and HRP conjugates facilitate the TMB oxidation in the presence of hydrogen peroxide, turning TMB into its blue oxidized product. ReadiUse™ TMB Substrate Solution is a premixed solution of TMB substrate with hydrogen peroxide. It produces a blue product upon interaction with HRP or HRP conjugates without the addition of hydrogen peroxide. The soluble blue product can be quantitated at 650 nm. Use of a stop solution enhances sensitivity 2-4 fold and the resulting yellow solution can be read at 450 nm. ReadiUse™ TMB Substrate Solution provides an convenient and ultrasensitive quantitative substrate system.
Example protocol
AT A GLANCE
Important
Warm ReadiUse™ TMB Solution to room temperature before use. Note The reagent is to be used as supplied, no dilution is required.
SAMPLE EXPERIMENTAL PROTOCOL
- Wash the assay plate following the incubation of HRP-labeled reagent.
- Add 100 µL of ReadiUse™ TMB Solution into each well.
- Incubate the plate at room temperature for 15 – 30 min or until the desired color develops. Note: The incubation time varies depending on the assay conditions.
- Measure the absorbance signal at 650 nm with an ELISA microplate reader. Note: If desired, the reaction can be stopped by adding an equal volume of 2M sulfuric acid to each well. Stopped reaction should be read at 450 nm.
Citations
View all 1 citations: Citation Explorer
Differential Effect of Chronic Morphine on Neuronal Degeneration in Male vs. Female Mice
Authors: Brazile Jr, Chet and Fan, Ruping and Benoit, Beau and Arnold, Thomas and Korneeva, Nadejda
Journal: Pathophysiology (2024): 152--165
Authors: Brazile Jr, Chet and Fan, Ruping and Benoit, Beau and Arnold, Thomas and Korneeva, Nadejda
Journal: Pathophysiology (2024): 152--165
References
View all 42 references: Citation Explorer
Au nanoparticles on citrate-functionalized graphene nanosheets with a high peroxidase-like performance
Authors: Chen X, Tian X, Su B, Huang Z, Oyama M.
Journal: Dalton Trans (2014): 7449
Authors: Chen X, Tian X, Su B, Huang Z, Oyama M.
Journal: Dalton Trans (2014): 7449
NiO nanoparticles modified with 5,10,15,20-tetrakis(4-carboxyl pheyl)-porphyrin: Promising peroxidase mimetics for HO and glucose detection
Authors: Liu Q, Yang Y, Li H, Zhu R, Shao Q, Yang S, Xu J.
Journal: Biosens Bioelectron (2014): 147
Authors: Liu Q, Yang Y, Li H, Zhu R, Shao Q, Yang S, Xu J.
Journal: Biosens Bioelectron (2014): 147
Interactions of macromolecular crowding agents and cosolutes with small-molecule substrates: effect on horseradish peroxidase activity with two different substrates
Authors: Aumiller WM, Jr., Davis BW, Hatzakis E, Keating CD.
Journal: J Phys Chem B (2014): 10624
Authors: Aumiller WM, Jr., Davis BW, Hatzakis E, Keating CD.
Journal: J Phys Chem B (2014): 10624
Biogenic magnetic nanoparticles from Burkholderia sp. YN01 exhibiting intrinsic peroxidase-like activity and their applications
Authors: Pan Y, Li N, Mu J, Zhou R, Xu Y, Cui D, Wang Y, Zhao M.
Journal: Appl Microbiol Biotechnol. (2014)
Authors: Pan Y, Li N, Mu J, Zhou R, Xu Y, Cui D, Wang Y, Zhao M.
Journal: Appl Microbiol Biotechnol. (2014)
5,10,15,20-tetrakis(4-carboxyl phenyl)porphyrin-CdS nanocomposites with intrinsic peroxidase-like activity for glucose colorimetric detection
Authors: Liu Q, Jia Q, Zhu R, Shao Q, Wang D, Cui P, Ge J.
Journal: Mater Sci Eng C Mater Biol Appl (2014): 177
Authors: Liu Q, Jia Q, Zhu R, Shao Q, Wang D, Cui P, Ge J.
Journal: Mater Sci Eng C Mater Biol Appl (2014): 177
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