Tetramethylbenzidine (TMB) - Ready to Use Chromogen - 100 ml
Tetramethylbenzidine (TMB) Ready-to Use Chromogen (Ready-to-use Peroxidase Chromogen Substrate for ELISA )
3-3’,5,5'-tetramethylbenzidine (TMB) is a soluble chromogen substrate for horseradish peroxidase detection systems. It is recommended for ELISA procedures. Biømeda Ready-to-Use TMB is a combination of buffer, chromogen and H2O2, as a substrate, in one convenient single component reagent. The use of TMB as a chromogen, when peroxidase-based immunochemical procedures are employed, provides the means of visualisation and measurement of analytical concentration or enzyme activity in quantitative assays.
TMB yields soluble blue end product that can be analyzed kinetically using the microtiter plate reader at wavelength of 650 nm. Reaction can be stopped by addition of 0.5 M sulfuric acid or 1.0 M Hydrochloric acid yielding in deep yellow color and read at 450 nm. When evaluating end point reaction visually, consider deep yellow color as positive and clear as negative.
TMB substrate is much more sensitive and less hazardous than benzidine or its other derivatives traditionally used as HRP chromogen substrates.
TMB is a ready to use chromogen for ELISA only. It is not suitable for immunohistochemistry.
Refrigerate at 4°C. Do not freeze.
Recommended Method: TMB Chromogen system is ready to use and requires no further preparations.
If the end point reaction is desired prepare 0.5 M solution of sulfuric acid or 0.5 M Hydrochloric acid. Sulfuric acid is not provided with the TMB reagent.
1.Following the ELISA Reagent containing the peroxidase, wash wells 3-4 times with 1X Immunoassay Buffer, Biømeda Cat. No. M61, or any other appropriate buffer. Remove air bubbles thoroughly.
2. Add 50 µl of Ready-to-Use TMB into each well Monitor color development until the positive control has the desired blue color strength. Color develops in 2-10 minutes at room temperature.
3. The reaction can be monitered as a function of time for kinetic assay or stopped with 0.1 % sodium fluoride and read at 650 nm.
4. If the procedure demands conversion to the yellow color, add 0.05 ml of 0.5 M sulfuric or HCL and record the absorbance at 450 nm.
Protect the TMB Chromogen from exposure to sunlight.
Do not use if reagent becomes turbid or blue color is present in the original container.
Discard unused reagent according to local and state regulations.
This is a laboratory reagent and should only be used by trained laboratory personnel.
Avoid contact with skin and clothes.-