Testing Data
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TMB Stable Liquid Substrate 1.56mM for HRPO Substrate
TMB Stable Liquid Substrate 1.56mM for HRPO (3,3',5,5'-Tetramethylbenzidine)
Applications
ELISA
Purity
Molecular Biology Grade
Synonyms
TMB Stable Liquid Substrate 1.56mM for HRPO; TMB Stable Liquid Substrate 1.56mM for HRPO (3; 3'; 5; 5'-Tetramethylbenzidine); TMB Stable Liquid Substrate 1.56mM for HRPO substrate
Purity/Purification
Molecular Biology Grade
Form/Format
Clear to pale yellow/orange
Applicable Applications for TMB Stable Liquid Substrate 1.56mM for HRPO substrate
ELISA (EL/EIA)
CAS Number
4827-17-7
pH
4.9 ± 0.03
Stability (18-26 degree C)
Passes test
Stability (4 degree C)
Passes test
Preparation and Storage
Stable for 18 months at RT. To extend the stability of product, refrigerate at 4 degree C.
Testing Data
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Testing Data
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Related Product Information for TMB Stable Liquid Substrate 1.56mM for HRPO substrate
TMB produces a soluble end product which is blue in color that is quantitated at 650nm. Upon addition of an equal volume of 0.5M/Liter HCl, the resulting yellow chromogen is quantitated at 450nm. The yellow product is stable at room temperature for at least 1 hour displaying less than a 1% loss in absorbance during this time interval. 1.56mM TMB is recommended for manual assays.
Benzidine and its derivatives are well documented electron donors for the horseradish peroxidase (HRP) /H2O2 oxidation/reduction system. These amines are known to be potent carcinogens.
The synthesis of TMB by Holland, et al., provided a reagent more sensitive than benzidine and much less hazardous due to ortho methylation.
Using H2O2 as substrate, the reaction of TMB with HRP proceeds through 2 phases. First, a charge transfer complex, the result of a 1-electron oxidation is formed. This is a blue product displaying an absorption maximum at 650nm. Further oxidation proceeds through a green color shift to the fully oxidized yellow diimine with an absorption maximum at 450nm.
The diimine is quantitatively formed only if the molar concentration of H2O2 is twice that of TMB. The molar extinction coefficient of the charge transfer complex is reported to be 3.9x10e4 mol-1 cm-1 and that of the diimine to be 5.9x10e4 mol-1 cm-1. The blue reaction product can be converted to the diimine non-enzymatically by the addition of acid providing an increase in sensitivity of at least 1.5.
1.56mM/Liter TMB is designed to measure low levels of HRP preventing the formation of the diimine allowing the user to determine enzyme activity or analyte concentration kinetically at 650nm or as an end point at 450nm after the addition of acid.
Benzidine and its derivatives are well documented electron donors for the horseradish peroxidase (HRP) /H2O2 oxidation/reduction system. These amines are known to be potent carcinogens.
The synthesis of TMB by Holland, et al., provided a reagent more sensitive than benzidine and much less hazardous due to ortho methylation.
Using H2O2 as substrate, the reaction of TMB with HRP proceeds through 2 phases. First, a charge transfer complex, the result of a 1-electron oxidation is formed. This is a blue product displaying an absorption maximum at 650nm. Further oxidation proceeds through a green color shift to the fully oxidized yellow diimine with an absorption maximum at 450nm.
The diimine is quantitatively formed only if the molar concentration of H2O2 is twice that of TMB. The molar extinction coefficient of the charge transfer complex is reported to be 3.9x10e4 mol-1 cm-1 and that of the diimine to be 5.9x10e4 mol-1 cm-1. The blue reaction product can be converted to the diimine non-enzymatically by the addition of acid providing an increase in sensitivity of at least 1.5.
1.56mM/Liter TMB is designed to measure low levels of HRP preventing the formation of the diimine allowing the user to determine enzyme activity or analyte concentration kinetically at 650nm or as an end point at 450nm after the addition of acid.
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Product Notes
The TMB Stable Liquid Substrate 1.56mM for HRPO (Catalog #AAA639241) is a Substrate and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's TMB Stable Liquid Substrate 1.56mM for HRPO can be used in a range of immunoassay formats including, but not limited to, ELISA (EL/EIA). Researchers should empirically determine the suitability of the TMB Stable Liquid Substrate 1.56mM for HRPO for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "TMB Stable Liquid Substrate 1.56mM for HRPO, Substrate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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