ReadiUse™ TMB Substrate Solution *Optimized for ELISA Assays with HRP Conjugates*
Ordering information
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Physical properties
Molecular weight | 240.34 |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
Storage | Refrigerated (2-8 °C); Minimize light exposure |
UNSPSC | 12171501 |
Related products
Overview | ![]() ![]() |
See also: Protein to Protein Conjugation, Biotin and Streptavidin, Enzyme-Linked Immunosorbent Assay (ELISA), Kinases, Horseradish Peroxidase (HRP) and Poly-HRP, Immunofluorescence, Oxidases, Secondary Antibodies, Serology Assays for Antibody Detection, Western Blotting Assays
CAS 54827-17-7 | Molecular weight 240.34 |
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.
Platform
Absorbance microplate reader
Absorbance | 650 nm |
Recommended plate | Clear bottom |
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.
Calculators
Common stock solution preparation
Table 1. Volume of appropriate solvent needed to reconstitute specific mass of ReadiUse™ TMB Substrate Solution *Optimized for ELISA Assays with HRP Conjugates* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 416.077 µL | 2.08 mL | 4.161 mL | 20.804 mL | 41.608 mL |
5 mM | 83.215 µL | 416.077 µL | 832.154 µL | 4.161 mL | 8.322 mL |
10 mM | 41.608 µL | 208.039 µL | 416.077 µL | 2.08 mL | 4.161 mL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Images
References
View all 42 references: Citation Explorer
Peroxidase-like activity of apoferritin paired gold clusters for glucose detection
Authors: Jiang X, Sun C, Guo Y, Nie G, Xu L.
Journal: Biosens Bioelectron (2014): 165
Authors: Jiang X, Sun C, Guo Y, Nie G, Xu L.
Journal: Biosens Bioelectron (2014): 165
Novel tungsten carbide nanorods: an intrinsic peroxidase mimetic with high activity and stability in aqueous and organic solvents
Authors: Li N, Yan Y, Xia BY, Wang JY, Wang X.
Journal: Biosens Bioelectron (2014): 521
Authors: Li N, Yan Y, Xia BY, Wang JY, Wang X.
Journal: Biosens Bioelectron (2014): 521
Glucose-sensitive colorimetric sensor based on peroxidase mimics activity of porphyrin-Fe3O4 nanocomposites
Authors: Liu Q, Li H, Zhao Q, Zhu R, Yang Y, Jia Q, Bian B, Zhuo L.
Journal: Mater Sci Eng C Mater Biol Appl (2014): 142
Authors: Liu Q, Li H, Zhao Q, Zhu R, Yang Y, Jia Q, Bian B, Zhuo L.
Journal: Mater Sci Eng C Mater Biol Appl (2014): 142
Peroxidase-Like Catalytic Activity of Ag3PO4 Nanocrystals Prepared by a Colloidal Route
Authors: Liu Y, Zhu G, Yang J, Yuan A, Shen X.
Journal: PLoS One (2014): e109158
Authors: Liu Y, Zhu G, Yang J, Yuan A, Shen X.
Journal: PLoS One (2014): e109158
Colorimetric estimation of human glucose level using gamma-Fe(2)O(3) nanoparticles: an easily recoverable effective mimic peroxidase
Authors: Mitra K, Ghosh AB, Sarkar A, Saha N, Dutta AK.
Journal: Biochem Biophys Res Commun (2014): 30
Authors: Mitra K, Ghosh AB, Sarkar A, Saha N, Dutta AK.
Journal: Biochem Biophys Res Commun (2014): 30
Visual detection of blood glucose based on peroxidase-like activity of WS2 nanosheets
Authors: Lin T, Zhong L, Song Z, Guo L, Wu H, Guo Q, Chen Y, Fu F, Chen G.
Journal: Biosens Bioelectron (2014): 302
Authors: Lin T, Zhong L, Song Z, Guo L, Wu H, Guo Q, Chen Y, Fu F, Chen G.
Journal: Biosens Bioelectron (2014): 302
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
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)
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
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
Application notes
A Meta-Analysis of Common Calcium Indicators
A New Protein Crosslinking Method for Labeling and Modifying Antibodies
A Novel Fluorescent Probe for Imaging and Detecting Hydroxyl Radical in Living Cells
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
Abbreviation of Common Chemical Compounds Related to Peptides
A New Protein Crosslinking Method for Labeling and Modifying Antibodies
A Novel Fluorescent Probe for Imaging and Detecting Hydroxyl Radical in Living Cells
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
Abbreviation of Common Chemical Compounds Related to Peptides